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Sample records for raising cre strategic

  1. Essential Not Optional: A Strategic Approach to Fund-Raising for Endowments

    ERIC Educational Resources Information Center

    Griswold, John S.; Jarvis, William F.

    2012-01-01

    This overview of the state of endowment giving comes at a time of urgent need in the nonprofit sector. With market returns uncertain, and spending restraint difficult, the moderate but measurable increase in donations in the previous year invites institutions to consider elevating fund-raising to a more strategic position within the organization.…

  2. How Strategic Presidential Leadership and Institutional Culture Influenced Fund-Raising Effectiveness at Spelman College

    ERIC Educational Resources Information Center

    Barrett, T. Gregory

    2006-01-01

    How have presidential leadership strategy, decision approaches, and institutional culture preferences influenced fund-raising effectiveness at a historically Black college for women? These conceptual dimensions guided a qualitative study that interviewed three recent Spelman College presidents and investigated documentary evidence to develop an…

  3. Raising Student Achievement in the Dayton Public Schools. Report of the Dayton Public Schools Strategic Support Teams.

    ERIC Educational Resources Information Center

    Council of the Great City Schools, Washington, DC.

    This report describes the Dayton Public Schools Strategic Support Teams (SSTs), which address stagnant student achievement among Dayton's students. They are composed of superintendents and other senior urban school managers from across the country who have struggled with the same issues as Dayton. Chapter 1 summarizes SST findings about challenges…

  4. 76 FR 38267 - The Federal Motor Carrier Safety Administration's 2011-2016 Strategic Plan: Raising the Safety Bar

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-29

    ... was held on September 8, 2010 (75 FR 53015, August 30, 2010). The new draft strategic plan, The... the comment, if submitted on behalf of an association, business, labor union, etc.). You may review... FR 3316) at http://edocket.access.gpo.gov/2008/pdf/E8-785.pdf . FOR FURTHER INFORMATION CONTACT:...

  5. Cre-mediated recombination in pituitary somatotropes

    PubMed Central

    Nasonkin, Igor O.; Potok, Mary Anne; Camper, Sally A.

    2009-01-01

    We report a transgenic line with highly penetrant cre recombinase activity in the somatotrope cells of the anterior pituitary gland. Expression of the cre transgene is under the control of the locus control region of the human growth hormone gene cluster and the rat growth hormone promoter. Cre recombinase activity was assessed with two different lacZ reporter genes that require excision of a floxed stop sequence for expression: a chick β-actin promoter with the CMV enhancer transgene and a ROSA26 knock-in. Cre activity is detectable in the developing pituitary after initiation of Gh transcription and persists through adulthood with high penetrance in Gh expressing cells and lower penetrance in lactotropes, a cell type that shares a common origin with somatotropes. This Gh-cre transgenic line is suitable for efficient, cell-specific deletion of floxed regions of genomic DNA in differentiated somatotropes and a subset of lactotrope cells of the anterior pituitary gland. PMID:19039787

  6. Cre-mediated recombination in pituitary somatotropes.

    PubMed

    Nasonkin, Igor O; Potok, Mary Anne; Camper, Sally A

    2009-01-01

    We report a transgenic line with highly penetrant cre recombinase activity in the somatotrope cells of the anterior pituitary gland. Expression of the cre transgene is under the control of the locus control region of the human growth hormone gene cluster and the rat growth hormone promoter. Cre recombinase activity was assessed with two different lacZ reporter genes that require excision of a floxed stop sequence for expression: a chick beta-actin promoter with the CMV enhancer transgene and a ROSA26 knock-in. Cre activity is detectable in the developing pituitary after initiation of Gh transcription and persists through adulthood with high penetrance in Gh expressing cells and lower penetrance in lactotropes, a cell type that shares a common origin with somatotropes. This Gh-cre transgenic line is suitable for efficient, cell-specific deletion of floxed regions of genomic DNA in differentiated somatotropes and a subset of lactotrope cells of the anterior pituitary gland. PMID:19039787

  7. Knockin of Cre Gene at Ins2 Locus Reveals No Cre Activity in Mouse Hypothalamic Neurons

    PubMed Central

    Li, Ling; Gao, Lin; Wang, Kejia; Ma, Xianhua; Chang, Xusheng; Shi, Jian-Hui; Zhang, Ye; Yin, Kai; Liu, Zhimin; Shi, Yuguang; Xie, Zhifang; Zhang, Weiping J.

    2016-01-01

    The recombination efficiency and cell specificity of Cre driver lines are critical for exploring pancreatic β cell biology with the Cre/LoxP approach. Some commonly used Cre lines are based on the short Ins2 promoter fragment and show recombination activity in hypothalamic neurons; however, whether this stems from endogenous Ins2 promoter activity remains controversial. In this study, we generated Ins2-Cre knockin mice with a targeted insertion of IRES-Cre at the Ins2 locus and demonstrated with a cell lineage tracing study that the Ins2 gene is not transcriptionally active in the hypothalamus. The Ins2-Cre driver line displayed robust Cre expression and activity in pancreatic β cells without significant alterations in insulin expression. In the brain, Cre activity was mainly restricted to the choroid plexus, without significant recombination detected in the hippocampus or hypothalamus by the LacZ or fluorescent tdTomato reporters. Furthermore, Ins2-Cre mice exhibited normal glucose tolerance and insulin secretion upon glucose stimulation in vivo. In conclusion, this Ins2-Cre driver line allowed high-fidelity detection of endogenous Ins2 promoter activity in vivo, and the negative activity in the hypothalamus demonstrated that this system is a promising alternative tool for studying β cell biology. PMID:26830324

  8. Postnatal Male Germ Cell Expression of Cre Recombinase in Tex101-iCre Transgenic Mice

    PubMed Central

    Lei, Zhenmin; Lin, Jing; Li, Xian; Li, Shengqiang; Zhou, Huaxin; Araki, Yoshihiko; Lan, Zi-Jian

    2010-01-01

    We have generated a transgenic mouse line which expresses improved Cre recombinase (iCre) under the control of the testis-expressed gene 101 (Tex101) promoter. This transgenic mouse line was named Tex101-iCre. Using the floxed ROSA reporter mice, we found that robust Cre recombinase activity was detected in postnatal testes with weak or no activity in other tissues. Within the testis, Cre recombinase was active in spermatogenic cells as early as the prospermatogonia stage at day 1 after birth. In 30- and 60-day-old mice, positive Cre recombinase activity was detected not only in prospermatogonia but also in spermatogenic cells at later stages of spermatogenesis. There was little or no Cre activity in interstitial cells. Breeding wild-type females with homozygous floxed fibroblast growth factor receptor 2 (Fgfr2) males carrying the Tex101-iCre transgene did not produce any progeny with the floxed Fgfr2 allele. All of the progeny inherited a recombined Fgfr2 allele, indicating that complete deletion of the floxed Fgfr2 allele can be achieved in the male germline by Tex101-iCre mice. Furthermore, FGFR2 protein was not detected in spermatocytes and spermatids of adult Fgfr2fl/fl;Tex101-iCre mice. Taken together, our results suggest that the Tex101-iCre mouse line allows the inactivation of a floxed gene in spermatogenic cells in adult mice, which will facilitate the functional characterization of genes in normal spermatogenesis and male fertility. PMID:20853429

  9. Fshb-iCre mice are efficient and specific Cre deleters for the gonadotrope lineage.

    PubMed

    Wang, Huizhen; Hastings, Richard; Miller, William L; Kumar, T Rajendra

    2016-01-01

    Genetic analysis of development and function of the gonadotrope cell lineage within mouse anterior pituitary has been greatly facilitated by at least three currently available Cre strains in which Cre was either knocked into the Gnrhr locus or expressed as a transgene from Cga and Lhb promoters. However, in each case there are some limitations including CRE expression in thyrotropes within pituitary or ectopic expression outside of pituitary, for example in some populations of neurons or gonads. Hence, these Cre strains often pose problems with regard to undesirable deletion of alleles in non-gonadotrope cells, fertility and germline transmission of mutant alleles. Here, we describe generation and characterization of a new Fshb-iCre deleter strain using 4.7 kb of ovine Fshb promoter regulatory sequences driving iCre expression exclusively in the gonadotrope lineage within anterior pituitary. Fshb-iCre mice develop normally, display no ectopic CRE expression in gonads and are fertile. When crossed onto a loxP recombination-mediated red to green color switch reporter mouse genetic background, in vivo CRE recombinase activity is detectable in gonadotropes at more than 95% efficiency and the GFP-tagged gonadotropes readily purified by fluorescence activated cell sorting. We demonstrate the applicability of this Fshb-iCre deleter strain in a mouse model in which Dicer is efficiently and selectively deleted in gonadotropes. We further show that loss of DICER-dependent miRNAs in gonadotropes leads to profound suppression of gonadotropins resulting in male and female infertility. Thus, Fshb-iCre mice serve as a new genetic tool to efficiently manipulate gonadotrope-specific gene expression in vivo. PMID:26472536

  10. Germline recombination in a novel Cre transgenic line, Prl3b1-Cre mouse.

    PubMed

    Al-Soudy, Al-Sayed; Nakanishi, Tsuyoshi; Mizuno, Seiya; Hasegawa, Yoshikazu; Shawki, Hossam H; Katoh, Megumi C; Basha, Walaa A; Ibrahim, Abdelaziz E; El-Shemy, Hany A; Iseki, Hiroyoshi; Yoshiki, Atsushi; Hiromori, Youhei; Nagase, Hisamitsu; Takahashi, Satoru; Oishi, Hisashi; Sugiyama, Fumihiro

    2016-07-01

    Spermatogenesis is a complex and highly regulated process by which spermatogonial stem cells differentiate into spermatozoa. To better understand the molecular mechanisms of the process, the Cre/loxP system has been widely utilized for conditional gene knockout in mice. In this study, we generated a transgenic mouse line that expresses Cre recombinase under the control of the 2.5 kbp of the Prolactin family 3, subfamily b, member 1 (Prl3b1) gene promoter (Prl3b1-cre). Prl3b1 was initially reported to code for placental lactogen 2 (PL-2) protein in placenta along with increased expression toward the end of pregnancy. PL-2 was found to be expressed in germ cells in the testis, especially in spermatocytes. To analyze the specificity and efficiency of Cre recombinase activity in Prl3b1-cre mice, the mice were mated with reporter R26GRR mice, which express GFP ubiquitously before and tdsRed exclusively after Cre recombination. The systemic examination of Prl3b1-cre;R26GRR mice revealed that tdsRed-positive cells were detected only in the testis and epididymis. Fluorescence imaging of Prl3b1-cre;R26GRR testes suggested that Cre-mediated recombination took place in the germ cells with approximately 74% efficiency determined by in vitro fertilization. In conclusion, our results suggest that the Prl3b1-cre mice line provides a unique resource to understand testicular germ-cell development. genesis 54:389-397, 2016. © 2016 Wiley Periodicals, Inc. PMID:27124574

  11. Phenotypic Characterization of MIP-CreERT1Lphi Mice With Transgene-Driven Islet Expression of Human Growth Hormone.

    PubMed

    Oropeza, Daniel; Jouvet, Nathalie; Budry, Lionel; Campbell, Jonathan E; Bouyakdan, Khalil; Lacombe, Julie; Perron, Gabrielle; Bergeron, Valerie; Neuman, Joshua C; Brar, Harpreet K; Fenske, Rachel J; Meunier, Clemence; Sczelecki, Sarah; Kimple, Michelle E; Drucker, Daniel J; Screaton, Robert A; Poitout, Vincent; Ferron, Mathieu; Alquier, Thierry; Estall, Jennifer L

    2015-11-01

    There is growing concern over confounding artifacts associated with β-cell-specific Cre-recombinase transgenic models, raising questions about their general usefulness in research. The inducible β-cell-specific transgenic (MIP-CreERT(1Lphi)) mouse was designed to circumvent many of these issues, and we investigated whether this tool effectively addressed concerns of ectopic expression and disruption of glucose metabolism. Recombinase activity was absent from the central nervous system using a reporter line and high-resolution microscopy. Despite increased pancreatic insulin content, MIP-CreERT mice on a chow diet exhibited normal ambient glycemia, glucose tolerance and insulin sensitivity, and appropriate insulin secretion in response to glucose in vivo and in vitro. However, MIP-CreERT mice on different genetic backgrounds were protected from high-fat/ streptozotocin (STZ)-induced hyperglycemia that was accompanied by increased insulin content and islet density. Ectopic human growth hormone (hGH) was highly expressed in MIP-CreERT islets independent of tamoxifen administration. Circulating insulin levels remained similar to wild-type controls, whereas STZ-associated increases in α-cell number and serum glucagon were significantly blunted in MIP-CreERT(1Lphi) mice, possibly due to paracrine effects of hGH-induced serotonin expression. These studies reveal important new insight into the strengths and limitations of the MIP-CreERT mouse line for β-cell research. PMID:26153246

  12. Evaluation of CRE-mediated excision approaches in Arabidopsis thaliana.

    PubMed

    Marjanac, Gordana; De Paepe, Annelies; Peck, Ingrid; Jacobs, Anni; De Buck, Sylvie; Depicker, Anna

    2008-04-01

    The ability of the CRE recombinase to catalyze excision of a DNA fragment flanked by directly repeated lox sites has been exploited to modify gene expression and proved to function well in particular case studies. However, very often variability in CRE expression and differences in efficiency of CRE-mediated recombination are observed. Here, various approaches were investigated to reproducibly obtain optimal CRE activity. CRE recombination was analyzed either by transforming the CRE T-DNA into plants containing a lox-flanked fragment or by transforming a T-DNA harboring a lox-flanked fragment into plants producing the CRE recombinase. Although somatic CRE-mediated excision of a lox-flanked fragment was obtained in all transformants, a variable amount of germline-transmitted deletions was found among different independent transformants, irrespective of the orientation of transformation. Also, the efficiency of CRE-mediated excision correlated well with the CRE mRNA level. In addition, CRE-mediated fragment excision was compared after floral dip and after root tissue transformation when transforming in a CRE-expressing background. Importantly, less CRE activity was needed to excise the lox-flanked fragment from the transferred T-DNA after root tissue transformation than after floral dip transformation. We hypothesize that this is correlated with the lower T-DNA copy number inserted during root transformation as compared to floral dip transformation. PMID:17541719

  13. Beyond knockouts: cre resources for conditional mutagenesis

    PubMed Central

    Murray, Stephen A.; Eppig, Janan T.; Smedley, Damian; Simpson, Elizabeth M.; Rosenthal, Nadia

    2013-01-01

    With the effort of the International Phenotyping Consortium (IMPC) to produce thousands of strains with conditional potential gathering steam, there is growing recognition that it must be supported by a rich toolbox of cre driver strains. The approaches to build cre strains have evolved in both sophistication and reliability, replacing first generation strains with tools that can target individual cell populations with incredible precision and specificity. The modest set of cre drivers generated by individual labs over the past 15+ years is now growing rapidly, thanks to a number of large-scale projects to produce new cre strains for the community. The power of this growing resource, however, depends upon the proper deep characterization of strain function, as even the best designed strain can display a variety of undesirable features that must be considered in experimental design. This must be coupled with the parallel development of informatics tools to provide functional data to the user, and facilitated access to the strains through public repositories. We will discuss the current progress on all of these fronts and the challenges that remain to ensure the scientific community can capitalize on the tremendous number of mouse resources at their disposal. PMID:22926223

  14. Photo-activatable Cre recombinase regulates gene expression in vivo.

    PubMed

    Schindler, Suzanne E; McCall, Jordan G; Yan, Ping; Hyrc, Krzystof L; Li, Mingjie; Tucker, Chandra L; Lee, Jin-Moo; Bruchas, Michael R; Diamond, Marc I

    2015-01-01

    Techniques allowing precise spatial and temporal control of gene expression in the brain are needed. Herein we describe optogenetic approaches using a photo-activatable Cre recombinase (PA-Cre) to stably modify gene expression in the mouse brain. Blue light illumination for 12 hours via optical fibers activated PA-Cre in the hippocampus, a deep brain structure. Two-photon illumination through a thinned skull window for 100 minutes activated PA-Cre within a sub-millimeter region of cortex. Light activation of PA-Cre may allow permanent gene modification with improved spatiotemporal precision compared to standard methods. PMID:26350769

  15. Cre-mediated site-specific recombination in zebrafish embryos.

    PubMed

    Thummel, Ryan; Burket, Christopher T; Brewer, Jeffrey L; Sarras, Michael P; Li, Li; Perry, Martin; McDermott, Jeffrey P; Sauer, Brian; Hyde, David R; Godwin, Alan R

    2005-08-01

    Cre-mediated site-specific recombination has become an invaluable tool for manipulation of the murine genome. The ability to conditionally activate gene expression or to generate chromosomal alterations with this same tool would greatly enhance zebrafish genetics. This study demonstrates that the HSP70 promoter can be used to inducibly control expression of an enhanced green fluorescent protein (EGFP) -Cre fusion protein. The EGFP-Cre fusion protein is capable of promoting recombination between lox sites in injected plasmids or in stably inherited transgenes as early as 2 hr post-heat shock induction. Finally, the levels of Cre expression achieved in a transgenic fish line carrying the HSP70-EGFP-cre transgene are compatible with viability and both male and female transgenic fish are fertile subsequent to induction of EGFP-Cre expression. Hence, our data suggests that Cre-mediated recombination is a viable means of manipulating gene expression in zebrafish. PMID:15977183

  16. A Study of Plazomicin Compared With Colistin in Patients With Infection Due to Carbapenem-Resistant Enterobacteriaceae (CRE)

    ClinicalTrials.gov

    2016-09-01

    Bloodstream Infections (BSI) Due to CRE; Hospital-Acquired Bacterial Pneumonia (HABP) Due to CRE; Ventilator-Associated Bacterial Pneumonia (VABP) Due to CRE; Complicated Urinary Tract Infection (cUTI) Due to CRE; Acute Pyelonephritis (AP) Due to CRE

  17. Identification of the CRE-1 Cellulolytic Regulon in Neurospora crassa

    PubMed Central

    Sun, Jianping; Glass, N. Louise

    2011-01-01

    Background In filamentous ascomycete fungi, the utilization of alternate carbon sources is influenced by the zinc finger transcription factor CreA/CRE-1, which encodes a carbon catabolite repressor protein homologous to Mig1 from Saccharomyces cerevisiae. In Neurospora crassa, deletion of cre-1 results in increased secretion of amylase and β-galactosidase. Methodology/Principal Findings Here we show that a strain carrying a deletion of cre-1 has increased cellulolytic activity and increased expression of cellulolytic genes during growth on crystalline cellulose (Avicel). Constitutive expression of cre-1 complements the phenotype of a N. crassa Δcre-1 strain grown on Avicel, and also results in stronger repression of cellulolytic protein secretion and enzyme activity. We determined the CRE-1 regulon by investigating the secretome and transcriptome of a Δcre-1 strain as compared to wild type when grown on Avicel versus minimal medium. Chromatin immunoprecipitation-PCR of putative target genes showed that CRE-1 binds to only some adjacent 5′-SYGGRG-3′ motifs, consistent with previous findings in other fungi, and suggests that unidentified additional regulatory factors affect CRE-1 binding to promoter regions. Characterization of 30 mutants containing deletions in genes whose expression level increased in a Δcre-1 strain under cellulolytic conditions identified novel genes that affect cellulase activity and protein secretion. Conclusions/Significance Our data provide comprehensive information on the CRE-1 regulon in N. crassa and contribute to deciphering the global role of carbon catabolite repression in filamentous ascomycete fungi during plant cell wall deconstruction. PMID:21980519

  18. Perinatal induction of Cre recombination with tamoxifen.

    PubMed

    Lizen, Benoit; Claus, Melissa; Jeannotte, Lucie; Rijli, Filippo M; Gofflot, Françoise

    2015-12-01

    Temporal control of site-specific recombination is commonly achieved by using a tamoxifen-inducible form of Cre or Flp recombinases. Although powerful protocols of induction have been developed for gene inactivation at adult stages or during embryonic development, induction of recombination at late gestational or early postnatal stages is still difficult to achieve. In this context, using the ubiquitous CMV-CreER(T2) transgenic mice, we have tested and validated two procedures to achieve recombination just before and just after birth. The efficiency of recombination was evaluated in the brain, which is known to be more problematic to target. For the late gestation treatment with tamoxifen, different protocols of complementary administration of progesterone and estrogen were tested. However, delayed delivery and/or mortality of pups due to difficult delivery were always observed. To circumvent this problem, pups were collected from tamoxifen-treated pregnant dams by caesarian section at E18.5 and given to foster mothers. For postnatal treatment, different dosages of tamoxifen were administered by intragastric injection to the pups during 3 or 4 days after birth. The efficiency of these treatments was analyzed at P7 using a transgenic reporter line. They were also validated with the Hoxa5 conditional allele. In conclusion, we have developed efficient procedures that allow achieving efficient recombination of floxed alleles at perinatal stages. These protocols will allow investigating the late/adult functions of many developmental genes, whose characterization has been so far restricted to embryonic development. PMID:26395370

  19. Postnatal male germ-cell expression of cre recombinase in Tex101-iCre transgenic mice.

    PubMed

    Lei, Zhenmin; Lin, Jing; Li, Xian; Li, Shengqiang; Zhou, Huaxin; Araki, Yoshihiko; Lan, Zi-Jian

    2010-12-01

    We have generated a transgenic mouse line that expresses improved Cre recombinase (iCre) under the control of the testis-expressed gene 101 (Tex101) promoter. This transgenic mouse line was named Tex101-iCre. Using the floxed ROSA reporter mice, we found that robust Cre recombinase activity was detected in postnatal testes with weak or no activity in other tissues. Within the testis, Cre recombinase was active in spermatogenic cells as early as the prospermatogonia stage at day 1 after birth. In 30- and 60-day-old mice, positive Cre recombinase activity was detected not only in prospermatogonia but also in spermatogenic cells at later stages of spermatogenesis. There was little or no Cre activity in interstitial cells. Breeding wild-type females with homozygous floxed fibroblast growth factor receptor 2 (Fgfr2) males carrying the Tex101-iCre transgene did not produce any progeny with the floxed Fgfr2 allele. All the progeny inherited a recombined Fgfr2 allele, indicating that complete deletion of the floxed Fgfr2 allele by Tex101-iCre can be achieved in the male germline. Furthermore, FGFR2 protein was not detected in spermatocytes and spermatids of adult Fgfr2(fl/fl) ;Tex101-iCre mice. Taken together, our results suggest that the Tex101-iCre mouse line allows the inactivation of a floxed gene in spermatogenic cells in adult mice, which will facilitate the functional characterization of genes in normal spermatogenesis and male fertility. PMID:20853429

  20. Expression and Functions of CreD, an Inner Membrane Protein in Stenotrophomonas maltophilia

    PubMed Central

    Huang, Hsin-Hui; Lin, Yi-Tsung; Chen, Wei-Ching; Huang, Yi-Wei; Chen, Shiang-Jiuun; Yang, Tsuey-Ching

    2015-01-01

    CreBC is a highly conserved two-component regulatory system (TCS) in several gram-negative bacteria, including Escherichia coli, Aeromonas spp., Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. CreD is a conserved gene that encodes a predicted inner-membrane protein and is located near the creBC loci. Activation of CreBC increases creD expression; therefore, creD expression is generally used as a measure of CreBC activation in E. coli, Aeromonas spp., and P. aeruginosa systems. In this article, we aim to elucidate the expression of creD and further to investigate its functions in S. maltophilia. In spite of a short intergenic region of 81 bp between creBC and creD, creD is expressed separately from the adjacent creBC operon and from a promoter immediately upstream of creD (PcreD) in S. maltophilia. We found that the promoter activity of PcreD is negatively regulated by the creBC TCS, positively regulated by the bacterial culture density, and not affected by β-lactams. Furthermore, creD expression is not significantly altered in the presence of the phosphor-mimic variant of CreB, CreB(D55E), which mimics activated CreB. The functions of CreD of S. maltophilia were assessed by comparison among the following: wild-type KJ; the creD isogenic mutant, KJΔCreD; and the complementary strain, KJΔCreD(pCreD). The mutant lacking creD had cell division defects and aberrations in cell envelope integrity, which then triggered the σE-mediated envelope stress response. Thus, the results indicated that CreD plays a critical role in the maintenance of envelope integrity. PMID:26698119

  1. Raise Your Profile: Build Your Program

    ERIC Educational Resources Information Center

    Nutefall, Jennifer E.; Gaspar, Deborah

    2008-01-01

    To raise the library's profile within the campus community, it is critical to create a strategic plan and align library goals with those of the university. At George Washington University's Gelman Library, the instruction librarians gained internal and external support to hire two new instruction librarians to better support collaboration with the…

  2. PVX-Cre-mediated marker gene elimination from transgenic plants.

    PubMed

    Kopertekh, L; Jüttner, G; Schiemann, J

    2004-07-01

    Cre recombinase gene from bacteriophage P1 was transiently expressed by a Potato Virus X (PVX)-based vector in transgenic lox -target Nicotiana benthamiana plants to remove the selectable marker gene. The target construct consisted of two directly oriented lox sites flanking a bar gene located between a gfp coding region and an upstream CaMV 35S promoter. The Cre-mediated excision of intervening sequence placed the gfp coding region under the transcriptional control of the CaMV 35S promoter. GFP activity was observed in PVX-Cre systemically infected leaves, regenerants from PVX-Cre infected explants and T1 progeny of these regenerants. PVX-Cre was removed efficiently from the regenerants by adding the nucleoside analogue ribavirin to the culture medium. Molecular data proved a correlation between gfp expression and precise site-specific excision of the bar gene in all examined transgenic lines. The frequency of recombination expressed as a percentage of regenerated plants exhibiting marker gene excision varied from 48% to 82%. These results demonstrate that a plant virus vector can be used efficiently to express cre recombinase in vivo providing an alternative method for the production of transgenic plants without marker genes. PMID:15604695

  3. Strategic Leadership

    ERIC Educational Resources Information Center

    Davies, Barbara; Davies, Brent

    2004-01-01

    This article explores the nature of strategic leadership and assesses whether a framework can be established to map the dimensions of strategic leadership. In particular it establishes a model which outlines both the organizational abilities and the individual characteristics of strategic leaders.

  4. Tissue Specific Expression of Cre in Rat Tyrosine Hydroxylase and Dopamine Active Transporter-Positive Neurons

    PubMed Central

    Liu, Zhenyi; Brown, Andrew; Fisher, Dan; Wu, Yumei; Warren, Joe; Cui, Xiaoxia

    2016-01-01

    The rat is a preferred model system over the mouse for neurological studies, and cell type-specific Cre expression in the rat enables precise ablation of gene function in neurons of interest, which is especially valuable for neurodegenerative disease modeling and optogenetics. Yet, few such Cre rats are available. Here we report the characterization of two Cre rats, tyrosine hydroxylase (TH)-Cre and dopamine active transporter (DAT or Slc6a3)-Cre, by using a combination of immunohistochemistry (IHC) and mRNA fluorescence in situ hybridization (FISH) as well as a fluorescent reporter for Cre activity. We detected Cre expression in expected neurons in both Cre lines. Interestingly, we also found that in Th-Cre rats, but not DAT-Cre rats, Cre is expressed in female germ cells, allowing germline excision of the floxed allele and hence the generation of whole-body knockout rats. In summary, our data demonstrate that targeted integration of Cre cassette lead to faithful recapitulation of expression pattern of the endogenous promoter, and mRNA FISH, in addition to IHC, is an effective method for the analysis of the spatiotemporal gene expression patterns in the rat brain, alleviating the dependence on high quality antibodies that are often not available against rat proteins. The Th-Cre and the DAT-Cre rat lines express Cre in selective subsets of dopaminergic neurons and should be particularly useful for researches on Parkinson’s disease. PMID:26886559

  5. Tissue Specific Expression of Cre in Rat Tyrosine Hydroxylase and Dopamine Active Transporter-Positive Neurons.

    PubMed

    Liu, Zhenyi; Brown, Andrew; Fisher, Dan; Wu, Yumei; Warren, Joe; Cui, Xiaoxia

    2016-01-01

    The rat is a preferred model system over the mouse for neurological studies, and cell type-specific Cre expression in the rat enables precise ablation of gene function in neurons of interest, which is especially valuable for neurodegenerative disease modeling and optogenetics. Yet, few such Cre rats are available. Here we report the characterization of two Cre rats, tyrosine hydroxylase (TH)-Cre and dopamine active transporter (DAT or Slc6a3)-Cre, by using a combination of immunohistochemistry (IHC) and mRNA fluorescence in situ hybridization (FISH) as well as a fluorescent reporter for Cre activity. We detected Cre expression in expected neurons in both Cre lines. Interestingly, we also found that in Th-Cre rats, but not DAT-Cre rats, Cre is expressed in female germ cells, allowing germline excision of the floxed allele and hence the generation of whole-body knockout rats. In summary, our data demonstrate that targeted integration of Cre cassette lead to faithful recapitulation of expression pattern of the endogenous promoter, and mRNA FISH, in addition to IHC, is an effective method for the analysis of the spatiotemporal gene expression patterns in the rat brain, alleviating the dependence on high quality antibodies that are often not available against rat proteins. The Th-Cre and the DAT-Cre rat lines express Cre in selective subsets of dopaminergic neurons and should be particularly useful for researches on Parkinson's disease. PMID:26886559

  6. Functional mapping of Cre recombinase by pentapeptide insertional mutagenesis.

    PubMed

    Petyuk, Vladislav; McDermott, Jeffrey; Cook, Malcolm; Sauer, Brian

    2004-08-27

    Cre is a site-specific recombinase from bacteriophage P1. It is a member of the tyrosine integrase family and catalyzes reciprocal recombination between specific 34-bp sites called loxP. To analyze the structure-function relationships of this enzyme, we performed large scale pentapeptide insertional mutagenesis to generate insertions of five amino acids at random positions in the protein. The high density of insertion mutations into Cre allowed us to identify an unexpected degree of functional tolerance to insertions into the 4-5 beta-hairpin and into the loop between helices J and K (both of which contact the DNA in the minor groove) and also into helix A. The phenotypes of the majority of inserts allowed us to confirm a variety of predictions made on the basis of sequence conservation, known three-dimensional structure, and proposed catalytic mechanism. In particular, most insertions into conserved regions or secondary structure elements inactivated Cre, and most insertions located in nonconserved, unstructured regions preserved Cre activity. Less expectedly, the non-conserved and poorly structured loops and linkers between helices A-B, E-F, and M-N did not tolerate insertions, thus identifying these as critical regions for recombinase activity. We purified and characterized in vitro several representatives of these "unexpected" Cre insertion mutants. The role of those regions in the recombination process is discussed. PMID:15218019

  7. Raising Children Who Refuse To Be Raised.

    ERIC Educational Resources Information Center

    Ziegler, Dave

    This book is about the struggle of raising children who work against your every effort as a parent or clinician to help them succeed in the world. Specifically, this book is about children who often do not respond to approaches offered in books and parent training programs for "normal" children. There are other books that address individual…

  8. Infection control implications of heterogeneous resistance mechanisms in carbapenem-resistant Enterobacteriaceae (CRE).

    PubMed

    Goodman, K E; Simner, P J; Tamma, P D; Milstone, A M

    2016-01-01

    The Centers for Disease Control and Prevention (CDC) defines carbapenem-resistant Enterobacteriaceae (CRE) based upon a phenotypic demonstration of carbapenem resistance. However, considerable heterogeneity exists within this definitional umbrella. CRE may mechanistically differ by whether they do or do not produce carbapenemases. Moreover, patients can acquire CRE through multiple pathways: endogenously through antibiotic selective pressure on intestinal microbiota, exogenously through horizontal transmission or through a combination of these factors. Some evidence suggests that non-carbapenemase-producing CRE may be more frequently acquired by antibiotic exposure and carbapenemase-producing CRE via horizontal transmission, but definitive data are lacking. This review examines types of CRE resistance mechanisms, antibiotic exposure and horizontal transmission pathways of CRE acquisition, and the implications of these heterogeneities to the development of evidence-based CRE healthcare epidemiology policies. In our Expert Commentary & Five-Year View, we outline specific nosocomial CRE knowledge gaps and potential methodological approaches for their resolution. PMID:26535959

  9. Mosaic pattern of Cre recombinase expression in cochlear outer hair cells of the Brn3.1 Cre mouse.

    PubMed

    Frenz, Silke; Rak, Kristen; Völker, Johannes; Jürgens, Lukas; Scherzad, Agmal; Schendzielorz, Philipp; Radeloff, Andreas; Jablonka, Sibylle; Hansen, Stefan; Mlynski, Robert; Hagen, Rudolf

    2015-04-15

    The Brn3.1 gene encodes for the protein Brn3.1, which is a member of the POU-IV class of transcription factors. Mutation leads to nonsyndromic human progressive hearing loss (DFNA15). To investigate the suitability of the Brn3.1 promoter for Cre recombinase-induced genetic recombination in cochlear hair cells, we established a transgenic Brn3.1 Cre mouse. This mouse line was crossbred with floxed ROSA26 and ROSA26 reporter mice. The cochleae were histologically analysed in cryosections at E16.5 and whole-mount preparations from P2 until P85. In addition, mice from all used strains and their recombinant offspring were tested electrophysiologically by auditory brainstem responses (ABR) and distorsion product otoacoustic emissions (DPOAE). Cre recombinase activity could be detected in P14 and P21 animals in a mosaic pattern in 26.3 and 9.9% of the outer hair cells, respectively. All investigated mice showed normal ABR and DPOAE values, indicating that neither insertion of the internal ribosome entry site (IRES) Cre cassette into the Brn3.1 gene led to abnormal auditory development nor did the reporter strains show inherited hearing disorders. This study shows that Cre expression under the control of the Brn3.1 promoter is feasible and that the insertion of the internal ribosome entry site Cre cassette into this locus exerted no effects on hearing development. Because of the inconstant pattern and the limited duration of expression, the application of the developed mouse line might be restricted. Also, the unchanged hearing capacity and structural integrity of the organ of Corti in available reporter lines indicate that they may be useful tools for hearing research. PMID:25714426

  10. Strategic weapons

    SciTech Connect

    Not Available

    1991-09-01

    This paper describes the Defense Department's process for formulating its strategic weapons targeting policy and translating that policy into a nuclear war plan-the Single Integrated Operational Plan. GAO provides information on the relationship between the strategic nuclear targeting process and the determination of requirements for nuclear weapons and related delivery systems, level of civilian oversight, and categories and types of targets. These strategic nuclear weapons systems, commonly known as the triad, include land-based intercontinental ballistic missiles, submarine-launched ballistic missiles, and strategic bombers armed with nuclear bombs and missiles.

  11. Raising Strong Daughters.

    ERIC Educational Resources Information Center

    Gadeberg, Jeanette

    In response to an alarming drop in girls' self-esteem in early adolescence, this parents' guide provides suggestions for raising daughters to become confident, healthy, and independent. Chapter 1, "Yesterday's Daughters," examines how cultural messages inhibit girls' development. Chapter 2, "Raising an Opinionated Daughter," suggests how to help…

  12. Raising and Transparency.

    ERIC Educational Resources Information Center

    Langacker, Ronald W.

    1995-01-01

    An account of the phenomena that transformational syntax handled by means of "raising" is formulated in the context of cognitive grammar. Raising is analyzed as a special case of the metonymy that relational expressions exhibit in regard to their choice of overtly coded arguments. The transparency of these constructions is explained. (83…

  13. Strategic Planning.

    ERIC Educational Resources Information Center

    Groff, Warren H.

    1983-01-01

    Reviews the strategic elements of an institutional plan: assessment of the external environment, auditing of institutional strengths and weaknesses, and matching of institutional strengths with external opportunities through the process of strategic goal setting. Urges community colleges to take action-oriented, dynamic, purposeful steps to shape…

  14. Unexpected severe consequences of Pikfyve deletion by aP2- or Aq-promoter-driven Cre expression for glucose homeostasis and mammary gland development.

    PubMed

    Ikonomov, Ognian C; Sbrissa, Diego; Delvecchio, Khortnal; A Rillema, James; Shisheva, Assia

    2016-06-01

    Systemic deficiency of PIKfyve, the evolutionarily conserved phosphoinositide kinase synthesizing cellular PtdIns5P and PtdIns(3,5)P2 and implicated in insulin signaling, causes early embryonic death in mice. In contrast, mice with muscle-specific Pikfyve disruption have normal lifespan but exhibit early-age whole-body glucose intolerance and muscle insulin resistance, thus establishing the key role of muscle PIKfyve in glucose homeostasis. Fat and muscle tissues control postprandial glucose clearance through different mechanisms, raising questions as to whether adipose Pikfyve disruption will also trigger whole-body metabolic abnormalities, and if so, what the mechanism might be. To clarify these issues, here we have characterized two new mouse models with adipose tissue disruption of Pikfyve through Cre recombinase expression driven by adipose-specific aP2- or adiponectin (Aq) promoters. Whereas both mouse lines were ostensibly normal until adulthood, their glucose homeostasis and systemic insulin sensitivity were severely dysregulated. These abnormalities stemmed in part from accelerated fat-cell lipolysis and elevated serum FFA Intriguingly, aP2-Cre-PIKfyve(fl/fl) but not Aq-Cre-PIKfyve(fl/fl) females had severely impaired pregnancy-induced mammary gland differentiation and lactogenesis, consistent with aP2-Cre-mediated Pikfyve excision in nonadipogenic tissues underlying this defect. Intriguingly, whereas mammary glands from postpartum control and Aq-Cre-PIKfyve(fl/fl) mice or ex vivo mammary gland explants showed profound upregulation of PIKfyve protein levels subsequent to prolactin receptor activation, such increases were not apparent in aP2-Cre-PIKfyve(fl/fl) females. Collectively, our data identify for the first time that adipose tissue Pikfyve plays a key role in the mechanisms regulating glucose homeostasis and that the PIKfyve pathway is critical in mammary epithelial differentiation during pregnancy and lactogenesis downstream of prolactin receptor

  15. Raising Consciousness about Sexism

    ERIC Educational Resources Information Center

    Delworth, Ursula

    1973-01-01

    The sexism consciousness raising exercises provided in this article can help both men and women understand how cultural attitudes toward the sexes are embedded in our individual psychological makeup. (Author)

  16. Fund Raising with Panache.

    ERIC Educational Resources Information Center

    Dedman, Robert

    1985-01-01

    The key to fund raising is the donor, and fund raisers should learn to cultivate potential donors, approach them with goals compatible with their own, supplement their thinking, get them involved, and swamp them with gratitude. (MSE)

  17. Raising the Titanic.

    ERIC Educational Resources Information Center

    Baker, Romona

    1990-01-01

    Described is an activity in which groups of students investigate engineering principles by writing a feasibility study to raise the luxury liner, Titanic. The problem statement and directions, and suggestions for problem solutions are included. (CW)

  18. Regulation of glycogen metabolism by the CRE-1, RCO-1 and RCM-1 proteins in Neurospora crassa. The role of CRE-1 as the central transcriptional regulator.

    PubMed

    Cupertino, Fernanda Barbosa; Virgilio, Stela; Freitas, Fernanda Zanolli; Candido, Thiago de Souza; Bertolini, Maria Célia

    2015-04-01

    The transcription factor CreA/Mig1/CRE-1 is a repressor protein that regulates the use of alternative carbon sources via a mechanism known as Carbon Catabolite Repression (CCR). In Saccharomyces cerevisiae, Mig1 recruits the complex Ssn6-Tup1, the Neurospora crassa RCM-1 and RCO-1 orthologous proteins, respectively, to bind to promoters of glucose-repressible genes. We have been studying the regulation of glycogen metabolism in N. crassa and the identification of the RCO-1 corepressor as a regulator led us to investigate the regulatory role of CRE-1 in this process. Glycogen content is misregulated in the rco-1(KO), rcm-1(RIP) and cre-1(KO) strains, and the glycogen synthase phosphorylation is decreased in all strains, showing that CRE-1, RCO-1 and RCM-1 proteins are involved in glycogen accumulation and in the regulation of GSN activity by phosphorylation. We also confirmed the regulatory role of CRE-1 in CCR and its nuclear localization under repressing condition in N. crassa. The expression of all glycogenic genes is misregulated in the cre-1(KO) strain, suggesting that CRE-1 also controls glycogen metabolism by regulating gene expression. The existence of a high number of the Aspergillus nidulans CreA motif (5'-SYGGRG-3') in the glycogenic gene promoters led us to analyze the binding of CRE-1 to some DNA motifs both in vitro by DNA gel shift and in vivo by ChIP-qPCR analysis. CRE-1 bound in vivo to all motifs analyzed demonstrating that it down-regulates glycogen metabolism by controlling gene expression and GSN phosphorylation. PMID:25889113

  19. Arity Raising in Manticore

    NASA Astrophysics Data System (ADS)

    Bergstrom, Lars; Reppy, John

    Compilers for polymorphic languages are required to treat values in programs in an abstract and generic way at the source level. The challenges of optimizing the boxing of raw values, flattening of argument tuples, and raising the arity of functions that handle complex structures to reduce memory usage are old ones, but take on newfound import with processors that have twice as many registers. We present a novel strategy that uses both control-flow and type information to provide an arity raising implementation addressing these problems. This strategy is conservative - no matter the execution path, the transformed program will not perform extra operations.

  20. Musashi1-CreERT2: A New Cre Line for Conditional Mutagenesis in Neural Stem Cells

    PubMed Central

    Takeda, Haruna; Koso, Hideto; Tessarollo, Lino; Copeland, Neal G.; Jenkins, Nancy A.

    2012-01-01

    The RNA-binding protein Musashi1 (Msi1) is one of two mammalian homologues of Drosophila Musashi, which is required for the asymmetric cell division of sensory organ precursor cells. In the mouse central nervous system (CNS) Msi1 is preferentially expressed in mitotically active progenitor cells in the ventricular zone (VZ) of the neural tube during embryonic development and in the subventricular zone (SVZ) of the postnatal brain. Previous studies showed that cells in the SVZ can contribute to long-term neurogenesis in the olfactory bulb (OB) but it remains unclear whether Msi1-expressing cells have self-renewing potential and can contribute to neurogenesis in the adult. Here we describe the generation of Msi1-CreERT2 knock-in mice and show by cell lineage tracing that Msi1-CreERT2-expressing cells mark neural stem cells (NSCs) in both the embryonic and adult brain. Msi1-CreERT2 mice thus represent a new tool in our arsenal for genetically manipulating NSCs, which will be essential for understanding the molecular mechanisms underlying neural development. PMID:23132814

  1. Raising Responsible Teenagers.

    ERIC Educational Resources Information Center

    Myers, Bob

    Some parents raise children without knowing anything about child development and are thus bewildered about how to deal with difficult behavior. This books addresses three important areas of parenting--child development, parenting style, and discipline--and tries to help parents gain the self-control needed to become competent adults. Chapters 1,…

  2. Raising Better Boys.

    ERIC Educational Resources Information Center

    Canada, Geoffrey

    2000-01-01

    The author of "Reaching Up For Manhood" discusses troubling social/environmental conditions confronting boys. Raising better boys requires caring adults, safer risk-taking situations, positive reinforcement, and role models. Parents should monitor boys' media exposure, provide moral education, broaden their cultural and natural-world experiences,…

  3. Hydrodynamic Delivery of Cre Protein to Lineage-Mark or Time-Stamp Mouse Hepatocytes In situ

    PubMed Central

    Sonsteng, Katherine M.; Prigge, Justin R.; Talago, Emily A.; June, Ronald K.; Schmidt, Edward E.

    2014-01-01

    Cre-responsive fluorescent marker alleles are powerful tools for cell lineage tracing in mice; however their utility is limited by regulation of Cre activity. When targeting hepatocytes, hydrodynamic delivery of a Cre-expression plasmid can convert Cre-responsive alleles without inducing the intracellular or systemic antiviral responses often associated with viral-derived Cre-expression vectors. In this method, rapid high-volume intravenous inoculation induces hepatocyte-targeted uptake of extracellular molecules. Here we tested whether hydrodynamic delivery of Cre protein or Cre fused to the HIV-TAT cell-penetrating peptide could convert Cre-responsive reporters in hepatocytes of mice. Hydrodynamic delivery of 2 nmol of either Cre or TAT-Cre protein converted the reporter allele in 5 to 20% of hepatocytes. Neither protein gave detectable Cre activity in endothelia, non-liver organs, or non-hepatocyte cells in liver. Using mice homozygous for a Cre-responsive marker that directs red- (Cre-naïve) or green- (Cre-converted) fluorescent proteins to the nucleus, we assessed sub-saturation Cre-activity. One month after hydrodynamic inoculation with Cre protein, 58% of hepatocyte nuclei that were green were also red, indicating that less than half of the hepatocytes that had obtained enough Cre to convert one marker allele to green were able to convert all alleles. For comparison, one month after hydrodynamic delivery of a Cre-expression plasmid with a weak promoter, only 26% of the green nuclei were also red. Our results show that hydrodynamic delivery of Cre protein allows rapid allelic conversion in hepatocytes, but Cre-activity is sub-saturating so many cells will not convert multiple Cre-responsive alleles. PMID:24626158

  4. Diverse Regulation of the CreA Carbon Catabolite Repressor in Aspergillus nidulans.

    PubMed

    Ries, Laure N A; Beattie, Sarah R; Espeso, Eduardo A; Cramer, Robert A; Goldman, Gustavo H

    2016-05-01

    Carbon catabolite repression (CCR) is a process that selects the energetically most favorable carbon source in an environment. CCR represses the use of less favorable carbon sources when a better source is available. Glucose is the preferential carbon source for most microorganisms because it is rapidly metabolized, generating quick energy for growth. In the filamentous fungus Aspergillus nidulans, CCR is mediated by the transcription factor CreA, a C2H2 finger domain DNA-binding protein. The aim of this work was to investigate the regulation of CreA and characterize its functionally distinct protein domains. CreA depends in part on de novo protein synthesis and is regulated in part by ubiquitination. CreC, the scaffold protein in the CreB-CreC deubiquitination (DUB) complex, is essential for CreA function and stability. Deletion of select protein domains in CreA resulted in persistent nuclear localization and target gene repression. A region in CreA conserved between Aspergillus spp. and Trichoderma reesei was identified as essential for growth on various carbon, nitrogen, and lipid sources. In addition, a role of CreA in amino acid transport and nitrogen assimilation was observed. Taken together, these results indicate previously unidentified functions of this important transcription factor. These novel functions serve as a basis for additional research in fungal carbon metabolism with the potential aim to improve fungal industrial applications. PMID:27017621

  5. Generation of CRISPR/Cas9-mediated bicistronic knock-in ins1-cre driver mice

    PubMed Central

    Hasegawa, Yoshikazu; Hoshino, Yoshikazu; Ibrahim, Abdelaziz E.; Kato, Kanako; Daitoku, Yoko; Tanimoto, Yoko; Ikeda, Yoshihisa; Oishi, Hisashi; Takahashi, Satoru; Yoshiki, Atsushi; Yagami, Ken-ichi; Iseki, Hiroyoshi; Mizuno, Seiya; Sugiyama, Fumihiro

    2016-01-01

    In the present study, we generated novel cre driver mice for gene manipulation in pancreatic β cells. Using the CRISPR/Cas9 system, stop codon sequences of Ins1 were targeted for insertion of cre, including 2A sequences. A founder of C57BL/6J-Ins1em1 (cre) Utr strain was produced from an oocyte injected with pX330 containing the sequences encoding gRNA and Cas9 and a DNA donor plasmid carrying 2A-cre. (R26GRR x C57BL/6J-Ins1em1 (cre) Utr) F1 mice were histologically characterized for cre-loxP recombination in the embryonic and adult stages; cre-loxP recombination was observed in all pancreatic islets examined in which almost all insulin-positive cells showed tdsRed fluorescence, suggesting β cell-specific recombination. Furthermore, there were no significant differences in results of glucose tolerance test among genotypes (homo/hetero/wild). Taken together, these observations indicated that C57BL/6J-Ins1em1 (cre) Utr is useful for studies of glucose metabolism and the strategy of bicistronic cre knock-in using the CRISPR/Cas9 system could be useful for production of cre driver mice. PMID:27053096

  6. Evidence for Baseline Retinal Pigment Epithelium Pathology in the Trp1-Cre Mouse

    PubMed Central

    Thanos, Aristomenis; Morizane, Yuki; Murakami, Yusuke; Giani, Andrea; Mantopoulos, Dimosthenis; Kayama, Maki; Roh, Mi In; Michaud, Norman; Pawlyk, Basil; Sandberg, Michael; Young, Lucy H.; Miller, Joan W.; Vavvas, Demetrios G.

    2012-01-01

    The increasing popularity of the Cre/loxP recombination system has led to the generation of numerous transgenic mouse lines in which Cre recombinase is expressed under the control of organ- or cell-specific promoters. Alterations in retinal pigment epithelium (RPE), a multifunctional cell monolayer that separates the retinal photoreceptors from the choroid, are prevalent in the pathogenesis of a number of ocular disorders, including age-related macular degeneration. To date, six transgenic mouse lines have been developed that target Cre to the RPE under the control of various gene promoters. However, multiple lines of evidence indicate that high levels of Cre expression can be toxic to mammalian cells. In this study, we report that in the Trp1-Cre mouse, a commonly used transgenic Cre strain for RPE gene function studies, Cre recombinase expression alone leads to RPE dysfunction and concomitant disorganization of RPE layer morphology, large areas of RPE atrophy, retinal photoreceptor dysfunction, and microglial cell activation in the affected areas. The phenotype described herein is similar to previously published reports of conditional gene knockouts that used the Trp1-Cre mouse, suggesting that Cre toxicity alone could account for some of the reported phenotypes and highlighting the importance of the inclusion of Cre-expressing mice as controls in conditional gene targeting studies. PMID:22429967

  7. System's flips in climate-related energy (CRE) systems

    NASA Astrophysics Data System (ADS)

    Ramos, Maria-Helena; Creutin, Jean-Dominique; Engeland, Kolbjørn; François, Baptiste; Renard, Benjamin

    2014-05-01

    Several modern environmental questions invite to explore the complex relationships between natural phenomena and human behaviour at a range of space and time scales. This usually involves a number of cause-effect (causal) relationships, linking actions and events. In lay terms, 'effect' can be defined as 'what happened' and 'cause', 'why something happened.' In a changing world or merely moving from one scale to another, shifts in perspective are expected, bringing some phenomena into the foreground and putting others to the background. Systems can thus flip from one set of causal structures to another in response to environmental perturbations and human innovations or behaviors, for instance, as space-time signatures are modified. The identification of these flips helps in better understanding and predicting how societies and stakeholders react to a shift in perspective. In this study, our motivation is to investigate possible consequences of the shift to a low carbon economy in terms of socio-technico systems' flips. The focus is on the regional production of Climate-Related Energy (CRE) (hydro-, wind- and solar-power). We search for information on historic shifts that may help defining the forcing conditions of abrupt changes and extreme situations. We identify and present a series of examples in which we try to distinguish the various tipping points, thresholds, breakpoints and regime shifts that are characteristic of complex systems in the CRE production domain. We expect that with these examples our comprehension of the question will be enriched, providing us the elements needed to better validate modeling attempts, to predict and manage flips of complex CRE production systems. The work presented is part of the FP7 project COMPLEX (Knowledge based climate mitigation systems for a low carbon economy; http://www.complex.ac.uk/).

  8. The Legacy of Nat Sternberg: The Genesis of Cre-lox Technology.

    PubMed

    Yarmolinsky, Michael; Hoess, Ronald

    2015-11-01

    Cre-lox of bacteriophage P1 has become one of the most widely used tools for genetic engineering in eukaryotes. The origins of this tool date to more than 30 years ago when Nat L. Sternberg discovered the recombinase, Cre, and its specific locus of crossover, lox, while studying the maintenance of bacteriophage P1 as a stable plasmid. Recombinations mediated by Cre assist in cyclization of the DNA of infecting phage and in resolution of prophage multimers created by generalized recombination. Early in vitro work demonstrated that, although it shares similarities with the well-characterized bacteriophage λ integration, Cre-lox is in many ways far simpler in its requirements for carrying out recombination. These features would prove critical for its development as a powerful and versatile tool in genetic engineering. We review the history of the discovery and characterization of Cre-lox and touch upon the present direction of Cre-lox research. PMID:26958905

  9. Strategic Planning.

    ERIC Educational Resources Information Center

    Vivelo, Frank Robert

    1992-01-01

    Describes the future environment facing community colleges, addressing the service population, demands for accountability and quality, and the need for currency. Identifies seven areas a strategic plan should address (e.g., mission, student success, instructional quality, resource development, diversity, operational efficiency, and community…

  10. Strategic Staffing

    ERIC Educational Resources Information Center

    Clark, Ann B.

    2012-01-01

    Business and industry leaders do not flinch at the idea of placing top talent in struggling departments and divisions. This is not always the case in public education. The Charlotte-Mecklenburg Schools made a bold statement to its community in its strategic plan by identifying two key reform levers--(1) an effective principal leading each school;…

  11. SInCRe-structural interactome computational resource for Mycobacterium tuberculosis.

    PubMed

    Metri, Rahul; Hariharaputran, Sridhar; Ramakrishnan, Gayatri; Anand, Praveen; Raghavender, Upadhyayula S; Ochoa-Montaño, Bernardo; Higueruelo, Alicia P; Sowdhamini, Ramanathan; Chandra, Nagasuma R; Blundell, Tom L; Srinivasan, Narayanaswamy

    2015-01-01

    We have developed an integrated database for Mycobacterium tuberculosis H37Rv (Mtb) that collates information on protein sequences, domain assignments, functional annotation and 3D structural information along with protein-protein and protein-small molecule interactions. SInCRe (Structural Interactome Computational Resource) is developed out of CamBan (Cambridge and Bangalore) collaboration. The motivation for development of this database is to provide an integrated platform to allow easily access and interpretation of data and results obtained by all the groups in CamBan in the field of Mtb informatics. In-house algorithms and databases developed independently by various academic groups in CamBan are used to generate Mtb-specific datasets and are integrated in this database to provide a structural dimension to studies on tuberculosis. The SInCRe database readily provides information on identification of functional domains, genome-scale modelling of structures of Mtb proteins and characterization of the small-molecule binding sites within Mtb. The resource also provides structure-based function annotation, information on small-molecule binders including FDA (Food and Drug Administration)-approved drugs, protein-protein interactions (PPIs) and natural compounds that bind to pathogen proteins potentially and result in weakening or elimination of host-pathogen protein-protein interactions. Together they provide prerequisites for identification of off-target binding. PMID:26130660

  12. Tamoxifen-Containing Eye Drops Successfully Trigger Cre-Mediated Recombination in the Entire Eye.

    PubMed

    Schlecht, Anja; Leimbeck, Sarah V; Tamm, Ernst R; Braunger, Barbara M

    2016-01-01

    Embryonic lethality in mice with targeted gene deletion is a major issue that can be circumvented by using Cre-loxP-based animal models. Various inducible Cre systems are available, e.g. such that are activated following tamoxifen treatment, and allow deletion of a specific target gene at any desired time point during the life span of the animal. In this study, we describe the efficiency of topical tamoxifen administration by eye drops using a Cre- reporter mouse strain (R26R). We report that tamoxifen-responsive CAGGCre-ER (TM) mice show a robust Cre- mediated recombination throughout the entire eye. PMID:26427451

  13. Efficient, specific, developmentally appropriate cre-mediated recombination in anterior pituitary gonadotropes and thyrotropes.

    PubMed

    Pérez-Millán, María Inés; Zeidler, Michael G; Saunders, Thomas L; Camper, Sally A; Davis, Shannon W

    2013-11-01

    Tissue-specific expression of cre recombinase is a well-established genetic tool to analyze gene function, and it is limited only by the efficiency and specificity of available cre mouse strains. Here, we report the generation of a transgenic line containing a cre cassette with codon usage optimized for mammalian cells (iCre) under the control of a mouse glycoprotein hormone α-subunit (αGSU) regulatory sequences in a bacterial artificial chromosome genomic clone. Initial analysis of this transgenic line, Tg(αGSU-iCre), with cre reporter strains reveals onset of cre activity in the differentiating cells of the developing anterior pituitary gland at embryonic day 12.5, with a pattern characteristic of endogenous αGSU. In adult mice, αGSU-iCre was active in the anterior lobe of the pituitary gland and in the cells that produce αGSU (gonadotropes and thyrotropes) with high penetrance. Little or no activity was observed in other tissues, including skeletal and cardiac muscle, brain, kidney, lungs, testis, ovary, and liver. This αGSU-iCre line is suitable for efficient, specific, and developmentally regulated deletion of floxed alleles in anterior pituitary gonadotropes and thyrotropes. PMID:23996951

  14. Pou4f2 knock-in Cre mouse: A multifaceted genetic tool for vision researchers

    PubMed Central

    Simmons, Aaron B.; Bloomsburg, Samuel J.; Billingslea, Samuel A.; Merrill, Morgan M.; Li, Shuai; Thomas, Marshall W.

    2016-01-01

    Purpose: A transgenic mouse that expresses Cre recombinase under control of the Pou4f2-promoter (also referred to as Brn-3b and Brn-3.2) was characterized. Pou4f2 expression has been reported in a subset of retinal ganglion cells (RGCs) in the retina, in the midbrain, and in the germline. In this study, we characterize the expression pattern of this Cre-recombinase line and report its utility in targeted deletion, temporal deletion, RGC depletion, and germline targeting, which can be regulated by the sex of the Cre-carrying mouse. Methods: Pou4f2Cre was mapped by using a combination of PCR and sequencing of PCR products to better understand the construct and to locate where it was inserted within the Pou4f2 locus. Cre expression patterns were examined by crossing Pou4f2Cre/+ mice to Cre reporter mice. Immunohistochemistry was used to further define the pattern of Cre expression and Cre-mediated recombination within the retina, brain, and other tissues. Results: An internal ribosome entry site (IRES)-Cre cassette was inserted into the Pou4f2 gene disrupting normal gene function, as verified by the depletion of RGCs in mice homozygous for the insert. Pou4f2Cre expression was observed in the retina, brain, peripheral neurons, and male germ cells. Germline recombination was observed when the sire carried the Cre and the target for recombination. In all other breeding schemes, recombination was observed within subsets of cells within the retina, brain, intestines, heart, and gonads. In the retina, Cre efficiently targets recombination in neurons within the RGC layer (RGL), the inner nuclear layer (INL), and a small percentage of photoreceptors, activity that has not been previously reported. Unlike most other Cre lines active in the inner retina, recombination in Müller and other glia was not observed in mice carrying Pou4f2Cre. Within the visual centers of the brain, Cre targets recombination in about 15% of cells within the superchiasmatic nucleus, lateral geniculate

  15. The CreC Regulator of Escherichia coli, a New Target for Metabolic Manipulations

    PubMed Central

    Godoy, Manuel S.; Nikel, Pablo I.; Cabrera Gomez, José G.

    2015-01-01

    The CreBC (carbon source-responsive) two-component regulation system of Escherichia coli affects a number of functions, including intermediary carbon catabolism. The impacts of different creC mutations (a ΔcreC mutant and a mutant carrying the constitutive creC510 allele) on bacterial physiology were analyzed in glucose cultures under three oxygen availability conditions. Differences in the amounts of extracellular metabolites produced were observed in the null mutant compared to the wild-type strain and the mutant carrying creC510 and shown to be affected by oxygen availability. The ΔcreC strain secreted more formate, succinate, and acetate but less lactate under low aeration. These metabolic changes were associated with differences in AckA and LdhA activities, both of which were affected by CreC. Measurement of the NAD(P)H/NAD(P)+ ratios showed that the creC510 strain had a more reduced intracellular redox state, while the opposite was observed for the ΔcreC mutant, particularly under intermediate oxygen availability conditions, indicating that CreC affects redox balance. The null mutant formed more succinate than the wild-type strain under both low aeration and no aeration. Overexpression of the genes encoding phosphoenolpyruvate carboxylase from E. coli and a NADH-forming formate dehydrogenase from Candida boidinii in the ΔcreC mutant further increased the yield of succinate on glucose. Interestingly, the elimination of ackA and adhE did not significantly improve the production of succinate. The diverse metabolic effects of this regulator on the central biochemical network of E. coli make it a good candidate for metabolic-engineering manipulations to enhance the formation of bioproducts, such as succinate. PMID:26497466

  16. Retinofugal Projections from Melanopsin-Expressing Retinal Ganglion Cells Revealed by Intraocular Injections of Cre-Dependent Virus

    PubMed Central

    Delwig, Anton; Larsen, DeLaine D.; Yasumura, Douglas; Yang, Cindy F.; Shah, Nirao M.; Copenhagen, David R.

    2016-01-01

    To understand visual functions mediated by intrinsically photosensitive melanopsin-expressing retinal ganglion cells (mRGCs), it is important to elucidate axonal projections from these cells into the brain. Initial studies reported that melanopsin is expressed only in retinal ganglion cells within the eye. However, recent studies in Opn4-Cre mice revealed Cre-mediated marker expression in multiple brain areas. These discoveries complicate the use of melanopsin-driven genetic labeling techniques to identify retinofugal projections specifically from mRGCs. To restrict labeling to mRGCs, we developed a recombinant adeno-associated virus (AAV) carrying a Cre-dependent reporter (human placental alkaline phosphatase) that was injected into the vitreous of Opn4-Cre mouse eyes. The labeling observed in the brain of these mice was necessarily restricted specifically to retinofugal projections from mRGCs in the injected eye. We found that mRGCs innervate multiple nuclei in the basal forebrain, hypothalamus, amygdala, thalamus and midbrain. Midline structures tended to be bilaterally innervated, whereas the lateral structures received mostly contralateral innervation. As validation of our approach, we found projection patterns largely corresponded with previously published results; however, we have also identified a few novel targets. Our discovery of projections to the central amygdala suggests a possible direct neural pathway for aversive responses to light in neonates. In addition, projections to the accessory optic system suggest that mRGCs play a direct role in visual tracking, responses that were previously attributed to other classes of retinal ganglion cells. Moreover, projections to the zona incerta raise the possibility that mRGCs could regulate visceral and sensory functions. However, additional studies are needed to investigate the actual photosensitivity of mRGCs that project to the different brain areas. Also, there is a concern of "overlabeling" with very

  17. Redesign of the monomer–monomer interface of Cre recombinase yields an obligate heterotetrameric complex

    PubMed Central

    Zhang, Chi; Myers, Connie A.; Qi, Zongtai; Mitra, Robi D.; Corbo, Joseph C.; Havranek, James J.

    2015-01-01

    Cre recombinase catalyzes the cleavage and religation of DNA at loxP sites. The enzyme is a homotetramer in its functional state, and the symmetry of the protein complex enforces a pseudo-palindromic symmetry upon the loxP sequence. The Cre-lox system is a powerful tool for many researchers. However, broader application of the system is limited by the fixed sequence preferences of Cre, which are determined by both the direct DNA contacts and the homotetrameric arrangement of the Cre monomers. As a first step toward achieving recombination at arbitrary asymmetric target sites, we have broken the symmetry of the Cre tetramer assembly. Using a combination of computational and rational protein design, we have engineered an alternative interface between Cre monomers that is functional yet incompatible with the wild-type interface. Wild-type and engineered interface halves can be mixed to create two distinct Cre mutants, neither of which are functional in isolation, but which can form an active heterotetramer when combined. When these distinct mutants possess different DNA specificities, control over complex assembly directly discourages recombination at unwanted half-site combinations, enhancing the specificity of asymmetric site recombination. The engineered Cre mutants exhibit this assembly pattern in a variety of contexts, including mammalian cells. PMID:26365240

  18. Generation and characterization of an endothelin-2 iCre mouse.

    PubMed

    Cacioppo, Joseph A; Koo, Yongbum; Lin, Po-Ching Patrick; Gal, Arnon; Ko, CheMyong

    2015-02-01

    A novel transgenic mouse line that expresses codon-improved Cre recombinase (iCre) under regulation of the Endothelin-2 gene (edn2) promoter was developed for the conditional deletion of genes in Endothelin-2 lineage cells and for the spatial and temporal localization of Endothelin-2 expression. Endothelin-2 (EDN2, ET-2, previously VIC) is a transcriptionally regulated 21 amino acid peptide implicated in vascular homeostasis, and more recently in female reproduction, gastrointestinal function, immunology, and cancer pathogenesis that acts through membrane receptors and G-protein signaling. A cassette (edn2-iCre) was constructed that contained iCre, a polyadenylation sequence, and a neomycin selection marker in front of the endogenous start codon of the edn2 gene in a mouse genome BAC clone. The cassette was introduced into the C57BL/6 genome by pronuclear injection, and two lines of edn2-iCre positive mice were produced. The edn2-iCre mice were bred with ROSA26-lacZ and Ai9 reporter mice to visualize areas of functional iCre expression. Strong expression was seen in the periovulatory ovary, stomach and small intestine, and colon. Uniquely, we report punctate expression in the corneal epithelium, the liver, the lung, the pituitary, the uterus, and the heart. In the embryo, expression is localized in developing hair follicles and the dermis. Therefore, edn2-iCre mice will serve as a novel line for conditional gene deletion in these tissues. PMID:25604013

  19. [The transgenic mice specifically express Cre recombinase in central nerve system].

    PubMed

    Sheng, Ji-Po; Hou, Ning; Cheng, Xuan; Yang, Xiao; Deng, Ji-Xian

    2004-12-01

    For specific expressing Cre recombinase in central nerve system (CNS), a transgenic construct (pGFAP-Cre-hGH), containing the beta-globin insulators, 1.8 kb of glial fibrillary acidic protein gene (GFAP) 5' end regulation region, Cre gene and polyA of human growth hormone gene (hGH) was generated, in which the 5' end regulation region of GFAP was isolated from a 129sv mouse genomic DNA library with PCR-screening. 7.6 kb of pGFAP-Cre-hGH DNA fragment was introduced into 191 fertilized eggs by microinjection. 176 injected eggs were implanted into the oviducts of eight female mice respectively, from which 25 offspring were obtained. Seven mice carry the Cre genes by the identification of PCR and Southern blotting, and the integration efficiency is 28%. The GFAP-Cre transgenic mice were crossed with ROSA26 mice whose genomic DNA is integrated by LoxP sites and LacZ expression frame to check the activity and the tissue-specific expression of Cre recombinase and recombination with its mediation between two LoxP sites. The results of LacZ dying showed that the Cre recombinase was expressed only in CNS and successfully mediated the recombination between the LoxP sites in vivo. PMID:15633637

  20. Generation and characterization of an Endothelin-2 iCre mouse

    PubMed Central

    Cacioppo, Joseph; Koo, Yongbum; Lin, Po-Ching Patrick; Gal, Arnon; Ko, CheMyong

    2015-01-01

    A novel transgenic mouse line that expresses codon-improved Cre recombinase (iCre) under regulation of the Endothelin-2 gene (edn2) promoter was developed for the conditional deletion of genes in Endothelin-2 lineage cells and for the spatial and temporal localization of Endothelin-2 expression. Endothelin-2 (EDN2, ET-2, previously VIC) is a transcriptionally regulated 21 amino acid peptide implicated in vascular homeostasis, and more recently in female reproduction, gastrointestinal function, immunology, and cancer pathogenesis that acts through membrane receptors and G-protein signaling. A cassette (edn2-iCre) was constructed that contained iCre, a polyadenylation sequence, and a neomycin selection marker in front of the endogenous start codon of the edn2 gene in a mouse genome BAC clone. The cassette was introduced into the C57BL/6 genome by pronuclear injection, and two lines of edn2-iCre positive mice were produced. The edn2-iCre mice were bred with ROSA26-lacZ and Ai9 reporter mice to visualize areas of functional iCre expression. Strong expression was seen in the periovulatory ovary, stomach and small intestine, and colon. Uniquely, we report punctate expression in the corneal epithelium, the liver, the lung, the pituitary, the uterus, and the heart. In the embryo, expression is localized in developing hair follicles and the dermis. Therefore, edn2-iCre mice will serve as a novel line for conditional gene deletion in these tissues. PMID:25604013

  1. Dre - Cre Sequential Recombination Provides New Tools for Retinal Ganglion Cell Labeling and Manipulation in Mice

    PubMed Central

    Shi, Melody; Liu, Pinghu; Dong, Lijin; Parmhans, Nadia; Popescu, Octavian; Badea, Tudor Constantin

    2014-01-01

    Background Genetic targeting methods have greatly advanced our understanding of many of the 20 Retinal Ganglion Cell (RGC) types conveying visual information from the eyes to the brain. However, the complexity and partial overlap of gene expression patterns in RGCs call for genetic intersectional or sparse labeling strategies. Loci carrying the Cre recombinase in conjunction with conditional knock-out, reporter or other genetic tools can be used for targeted cell type ablation and functional manipulation of specific cell populations. The three members of the Pou4f family of transcription factors, Brn3a, Brn3b and Brn3c, expressed early during RGC development and in combinatorial pattern amongst RGC types are excellent candidates for such gene manipulations. Methods and Findings We generated conditional Cre knock-in alleles at the Brn3a and Brn3b loci, Brn3aCKOCre and Brn3bCKOCre. When crossed to mice expressing the Dre recombinase, the endogenous Brn3 gene expressed by Brn3aCKOCre or Brn3bCKOCre is removed and replaced with a Cre recombinase, generating Brn3aCre and Brn3bCre knock-in alleles. Surprisingly both Brn3aCre and Brn3bCre knock-in alleles induce early ubiquitous recombination, consistent with germline expression. However in later stages of development, their expression is limited to the expected endogenous pattern of the Brn3a and Brn3b genes. We use the Brn3aCre and Brn3bCre alleles to target a Cre dependent Adeno Associated Virus (AAV) reporter to RGCs and demonstrate its use in morphological characterization, early postnatal gene delivery and tracing the expression of Brn3 genes in RGCs. Conclusions Dre recombinase effectively recombines the Brn3aCKOCre and Brn3bCKOCre alleles containing its roxP target sites. Sequential Dre to Cre recombination reveals Brn3a and Brn3b expression in early mouse development. The generated Brn3aCre and Brn3bCre alleles are useful tools that can target exogenously delivered Cre dependent reagents to RGCs in early

  2. Germ cell-specific expression of Cre recombinase using the VASA promoter in the pig.

    PubMed

    Song, Yuning; Lai, Liangxue; Li, Li; Huang, Yongye; Wang, Anfeng; Tang, Xiaochun; Pang, Daxin; Li, Zhanjun; Ouyang, Hongsheng

    2016-01-01

    The Cre-loxP system is a powerful tool for genetic analysis of distinct cell lineages and tissue-specific gene knockout in animal models. VASA is specifically expressed in reproductive tissues, and is known to play important roles in spermatogenesis and germ-cell growth. In this study, Cre recombinase transgenic pigs under the control of the VASA promoter were generated by somatic cell nuclear transfer. Germ cell-specific expression of Cre recombinase in VASA-Cre transgenic pigs was shown by western blotting and immunohistochemistry. VASA-Cre transgenic pigs will be a useful tool for germ cell-specific gene knockout and a disease model for disorders of the reproductive system. PMID:27047735

  3. Systolic dysfunction in cardiac-specific ligand-inducible MerCreMer transgenic mice

    PubMed Central

    Hall, Michael E.; Smith, Grant; Hall, John E.

    2011-01-01

    The Cre-loxP system is a useful tool to study the physiological effects of gene knockout in the heart. One limitation with using this system in the heart is the toxic effect of chronic expression of the Cre recombinase. To circumvent this limitation, a widely used inducible cardiac-specific model, Myh6-MerCreMer (Cre), using tamoxifen (TAM) to activate Cre has been developed. The current study examined cardiac function in Cre-positive C57B/J6 mice exposed to one, three, or five daily doses of a 40 mg/kg TAM to induce Cre activity specifically in the heart. Echocardiography demonstrated no statistically significant differences in systolic function (SF) at baseline as assessed by fractional shortening. In mice exposed to five injections, a significant fall in all determinants of SF was observed 6 days after TAM was initiated. However, SF returned to baseline levels 10 days after TAM initiation although the hearts exhibited significant hypertrophy. Heart weight-to-tibia length ratios were 73 ± 3, 78.5 ± 6, and 87.6 ± 9 mg/cm for one, three, and five TAM injections, respectively. TAM had no effect on cardiac function or hypertrophy in Cre-negative mice. Cre-positive mice receiving five TAM injections had significant reductions in cardiac mitochondrial ATP and significant reductions in the expression of proteins important for the regulation of cardiac oxidative phosphorylation including peroxisome proliferator-activated receptor-γ coactivator-1α and pyruvate dehydrogenase kinase-4. Thus inducible cardiac-specific activation of Cre recombinase caused a transient decline in SF that was dependent on the number of TAM doses and associated with significant hypertrophy and alterations in mitochondrial ATP and important proteins involved in the regulation of cardiac oxidative phosphorylation. PMID:21536850

  4. On Strategic Management: A Conversation with George Wilkinson.

    ERIC Educational Resources Information Center

    Brandt, Ron

    1991-01-01

    George Wilkinson, who has managed planning for United Way of America since 1980, observes that local planners using strategic planning raise more money and solve more community problems than those without such a process. Marketing is identical to strategic planning; both involve looking at an organization, doing and analysis, examining the future,…

  5. A New Strategic Planning Model for Academic Libraries.

    ERIC Educational Resources Information Center

    Birdsall, Douglas G.; Hensley, Oliver D.

    1994-01-01

    Presents a strategic planning model originally developed for university research and illustrates its usefulness for academic libraries. Library development and fund-raising activities are used as examples to illustrate the various stages of strategic planning, including scanning the environment, designing the plan, acceptance, and adoption.…

  6. Isolation of Novel CreERT2-Driver Lines in Zebrafish Using an Unbiased Gene Trap Approach.

    PubMed

    Jungke, Peggy; Hammer, Juliane; Hans, Stefan; Brand, Michael

    2015-01-01

    Gene manipulation using the Cre/loxP-recombinase system has been successfully employed in zebrafish to study gene functions and lineage relationships. Recently, gene trapping approaches have been applied to produce large collections of transgenic fish expressing conditional alleles in various tissues. However, the limited number of available cell- and tissue-specific Cre/CreERT2-driver lines still constrains widespread application in this model organism. To enlarge the pool of existing CreERT2-driver lines, we performed a genome-wide gene trap screen using a Tol2-based mCherry-T2a-CreERT2 (mCT2aC) gene trap vector. This cassette consists of a splice acceptor and a mCherry-tagged variant of CreERT2 which enables simultaneous labeling of the trapping event, as well as CreERT2 expression from the endogenous promoter. Using this strategy, we generated 27 novel functional CreERT2-driver lines expressing in a cell- and tissue-specific manner during development and adulthood. This study summarizes the analysis of the generated CreERT2-driver lines with respect to functionality, expression, integration, as well as associated phenotypes. Our results significantly enlarge the existing pool of CreERT2-driver lines in zebrafish and combined with Cre-dependent effector lines, the new CreERT2-driver lines will be important tools to manipulate the zebrafish genome. PMID:26083735

  7. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain

    PubMed Central

    Madisen, Linda; Zwingman, Theresa A.; Sunkin, Susan M.; Oh, Seung Wook; Zariwala, Hatim A.; Gu, Hong; Ng, Lydia L.; Palmiter, Richard D.; Hawrylycz, Michael J.; Jones, Allan R.; Lein, Ed S.; Zeng, Hongkui

    2009-01-01

    The Cre/lox system is widely used in mice to achieve cell-type-specific gene expression. However, a strong and universal responding system to express genes under Cre control is still lacking. We have generated a set of Cre reporter mice with strong, ubiquitous expression of fluorescent proteins of different spectra. The robust native fluorescence of these reporters enables direct visualization of fine dendritic structures and axonal projections of the labeled neurons, which is useful in mapping neuronal circuitry, imaging and tracking specific cell populations in vivo. Using these reporters and a high-throughput in situ hybridization platform, we are systematically profiling Cre-directed gene expression throughout the mouse brain in a number of Cre-driver lines, including novel Cre lines targeting different cell types in the cortex. Our expression data are displayed in a public online database to help researchers assess the utility of various Cre-driver lines for cell-type-specific genetic manipulation. PMID:20023653

  8. Cardiac-specific activation of Cre expression at late fetal development

    SciTech Connect

    Opherk, Jan P.; Yampolsky, Peter; Hardt, Stefan E.; Schoels, Wolfgang; Katus, Hugo A.; Koenen, Michael . E-mail: koenen@mpimf-heidelberg.mpg.de; Zehelein, Joerg

    2007-07-27

    In a first step towards dissecting molecular mechanisms that contribute to the development of cardiac diseases, we have generated transgenic mice that express a Cre-GFP fusion protein under the transcriptional control of a 4.3 kb murine cardiac Troponin I gene (cTnI) promoter. Cre-GFP expression, similar in three transgenic lines, is described in one line. In mouse embryos, transgenic for the Cre-GFP and ROSA lacZ reporter allele, first Cre-mediated recombination appeared at 16.5 dpc selectively at the heart. Like the endogenous cTnI gene, transgenic Cre expression showed a slow rise through fetal development that increased neonatally. Bitransgenic hearts, stained at 30 days of age, showed intense signals in ventricular and atrial myocytes while no recombination occurred in other tissues. The delayed onset of Cre activity in cTnI-Cre mice could provide a useful genetic tool to evaluate the function of loxP targeted cardiac genes without interference of recombination during early heart development.

  9. Advanced zebrafish transgenesis with Tol2 and application for Cre/lox recombination experiments.

    PubMed

    Mosimann, Christian; Zon, Leonard I

    2011-01-01

    Spatio-temporal transgene regulation by transgenic DNA recombinases is a central tool for genetic research in multicellular organisms, with excellent applications for misexpression and lineage tracing experiments. Cre recombinase-controlled lox site recombination is a cornerstone of contemporary mouse genetics, and Cre/lox techniques therefore attract increasing interest in the zebrafish field. Tol2-mediated zebrafish transgenesis now provides a stable platform for lox cassette transgenes, while the ease of drug treatments in zebrafish makes the model an ideal candidate for Tamoxifen/4-hydroxytamoxifen-inducible CreER(T2) experiments. In this chapter, we will first introduce the basics of Cre/lox methodology, CreER(T2) regulation by Tamoxifen/4-hydroxytamoxifen, as well as the benefits of Tol2 transgenesis for Cre/lox experiments. We will then in detail outline practical experimental steps for Tol2 transgenesis toward the creation of single-insertion transgenes. Lastly, we will introduce protocols for 4-hydroxytamoxifen-mediated CreER(T2) induction to perform spatio-temporal lox transgene regulation experiments in zebrafish embryos. PMID:21924163

  10. Heat-inducible Cre-lox system for marker excision in transgenic rice.

    PubMed

    Khattri, Abhilasha; Nandy, Soumen; Srivastava, Vibha

    2011-03-01

    The present study assessed the efficacy of a heat-inducible cre gene for conditional removal of the marker gene from a rice genome via Cre-lox recombination. A cre gene controlled by the soybean heat-shock promoter was introduced into the rice genome along with the recombination target (lox) construct. Cre-mediated recombination was expected to remove the marker gene and activate the promoter-less GUS gene. Six transgenic lines displayed well-regulated heat-inducible Cre activity in the callus. However, only one line that contained a single copy of the cre gene maintained this property in the regenerated plants and their progeny. Marker-free progeny were obtained from the plant that was heat-treated at the seedling stage, indicating the inheritance of the recombination 'footprint'. The presence of the 'footprint' was verified by polymerase chain reaction and Southern analysis. Therefore, the cre gene controlled by the soybean heat-shock promoter is an effective tool for conditional removal of the marker gene in rice. PMID:21451246

  11. Reversion to wildtype of a mutated and nonfunctional coxsackievirus B3CRE(2C).

    PubMed

    Smithee, Shane; Tracy, Steven; Chapman, Nora M

    2016-07-15

    The cis-acting replication element (CRE) in the 2C protein coding region [CRE(2C)] of enteroviruses (EV) facilitates the addition of two uridine residues (uridylylation) onto the virus-encoded protein VPg in order for it to serve as the RNA replication primer. We demonstrated that coxsackievirus B3 (CVB3) is replication competent in the absence of a native (uridylylating) CRE(2C) and also demonstrated that lack of a functional CRE(2C) led to generation of 5' terminal genomic deletions in the CVB3 CRE-knock-out (CVB3-CKO) population. We asked whether reversion of the mutated CRE(2C) occurred, thus permitting sustained replication, and when were 5' terminal deletions generated during replication. Virions were isolated from HeLa cells previously electroporated with infectious CVB3-CKO T7 transcribed RNA or from hearts and spleens of mice after transfection with CVB3-CKO RNA. Viral RNA was isolated in order to amplify the CRE(2C) coding region and the genomic 5' terminal sequences. Sequence analysis revealed reversion of the CVB3-CKO sequence to wildtype occurs by 8 days post-electroporation of HeLa cells and by 20days post-transfection in mice. However, 5' terminal deletions evolve prior to these times. Reversion of the CRE(2C) mutations to wildtype despite loss of the genomic 5' termini is consistent with the hypothesis that an intact CRE(2C) is inherently vital to EV replication even when it is not enabling efficient positive strand initiation. PMID:27130630

  12. Profiling of functional phosphodiesterase in mesangial cells using a CRE-SEAP-based reporting system

    PubMed Central

    Zhu, Ying; Yao, Jian; Meng, Yiman; Kasai, Ayumi; Hiramatsu, Nobuhiko; Hayakawa, Kunihiro; Miida, Takashi; Takeda, Masayuki; Okada, Masahiko; Kitamura, Masanori

    2006-01-01

    Phosphodiesterases (PDEs) are critically implicated in the regulation of mesangial cell function, but profile of functional PDEs in mesangial cells is still unclear. In this study, we investigated roles of individual PDEs in the regulation of mesangial cell behavior by the cAMP pathway. Reporter mesangial cells that express secreted alkaline phosphatase (SEAP) under the control of the cAMP response element (CRE) were exposed to selective PDE inhibitors in the presence or absence of cAMP, and activity of CRE, expression of CRE-regulated protein, mitogenesis and cell survival were examined. Exposure of reporter cells to cAMP-elevating agents resulted in time- and concentration-dependent activation of CRE. Treatment of the cells with any PDE inhibitors alone did not induce CRE activation. Under stimulation with 8-bromo-cAMP or 8-bromo-cGMP, however, inhibitors of PDE2, PDE3, PDE4 and PDE5 enhanced activation of CRE. Inhibition of PDE1 or PDE6 did not affect the CRE activation. Among different combinations tested, only inhibitors of PDE3 and PDE4 cooperatively increased the level of intracellular cAMP, activity of protein kinase A, activation of CRE, and CRE-regulated protein, connexin43. Concomitant inhibition of PDE3 and PDE4 attenuated mitogen-induced activation of extracellular signal-regulated kinases and cell proliferation. Under serum deprivation, combinational inhibition of PDE3 and PDE4 exclusively caused activation of caspase-3 and apoptosis. The present data elucidated that PDE3 and PDE4 play critical roles in the regulation of mesangial cell function. PDE3 and PDE4 were identified as the novel, antiapoptotic machinery that supports survival of mesangial cells. PMID:16751794

  13. Targeted DNA recombination in vivo using an adenovirus carrying the cre recombinase gene.

    PubMed Central

    Wang, Y; Krushel, L A; Edelman, G M

    1996-01-01

    Conditional gene expression and gene deletion are important experimental approaches for examining the functions of particular gene products in development and disease. The cre-loxP system from bacteriophage P1 has been used in transgenic animals to induce site-specific DNA recombination leading to gene activation or deletion. To regulate the recombination in a spatiotemporally controlled manner, we constructed a recombinant adenoviral vector, Adv/cre, that contained the cre recombinase gene under regulation of the herpes simplex virus thymidine kinase promoter. The efficacy and target specificity of this vector in mediating loxP-dependent recombination were analyzed in mice that had been genetically engineered to contain loxP sites in their genome. After intravenous injection of the Adv/cre vector into adult animals, the liver and spleen showed the highest infectivity of the adenovirus as well as the highest levels of recombination, whereas other tissues such as kidney, lung, and heart had lower levels of infection and recombination. Only trace levels of recombination were detected in the brain. However, when the Adv/cre vector was injected directly into specific regions of the adult brain, including the cerebral cortex, hippocampus, and cerebellum, recombination was detectable at the injection site. Furthermore, when the Adv/cre vector was injected into the forebrains of neonatal mice, the rearranged toxP locus from recombination could be detected in the injected regions for at least 8 weeks. Taken together, these results demonstrate that the Adv/cre vector expressing a functional cre protein is capable of mediating loxP-dependent recombination in various tissues and the recombined gene locus may in some cases be maintained for an extended period. The use of the adenovirus vector expressing cre combined with localized delivery to specific tissues may provide an efficient means to achieve conditional gene expression or knockout with precise spatiotemporal control

  14. Strategic Planning: What's so Strategic about It?

    ERIC Educational Resources Information Center

    Strong, Bart

    2005-01-01

    The words "strategic" and "planning" used together can lead to confusion unless one spent the early years of his career in never-ending, team-oriented, corporate training sessions. Doesn't "strategic" have something to do with extremely accurate bombing or a defensive missile system or Star Wars or something? Don't "strategic" and "planning" both…

  15. Pancreas-specific Cre driver lines and considerations for their prudent use

    PubMed Central

    Magnuson, Mark A.; Osipovich, Anna B.

    2013-01-01

    Summary Cre/LoxP has broad utility for studying the function, development and oncogenic transformation of pancreatic cells in mice. Here we provide an overview of the many different Cre driver lines that are available for such studies. We, discuss how variegated expression, transgene silencing, and recombination in undesired cell types have conspired to limit the performance of these lines sometimes leading to serious experimental concerns. We also discuss. preferred strategies for achieving high fidelity driver lines and remind investigators of the continuing need for caution when interpreting results obtained from any Cre/LoxP-based experiment performed in mice. PMID:23823474

  16. Effect of deletion mutation on the recombination activity of Cre recombinase.

    PubMed

    Rongrong, Liu; Lixia, Wang; Zhongping, Lin

    2005-01-01

    Cre recombinase from bacteriophage P1 is widely used in both in vitro and in vivo DNA manipulations. Based on a structural and functional analysis, three deleted cre mutants were constructed and expressed in Escherichia coli. Mutated recombinases were purified and their recombination activities were determined in vitro. Our results revealed that the mutant with amino-terminal deletion retains the recombination activity as high as wild type Cre; however, the carboxy-terminal deletion and the middle region deletion both lead to a complete loss of the recombinase function. PMID:15912212

  17. The Notch intracellular domain represses CRE-dependent transcription.

    PubMed

    Hallaq, Rania; Volpicelli, Floriana; Cuchillo-Ibanez, Inmaculada; Hooper, Claudie; Mizuno, Keiko; Uwanogho, Dafe; Causevic, Mirsada; Asuni, Ayodeji; To, Alvina; Soriano, Salvador; Giese, K Peter; Lovestone, Simon; Killick, Richard

    2015-03-01

    Members of the cyclic-AMP response-element binding protein (CREB) transcription factor family regulate the expression of genes needed for long-term memory formation. Loss of Notch impairs long-term, but not short-term, memory in flies and mammals. We investigated if the Notch-1 (N1) exerts an effect on CREB-dependent gene transcription. We observed that N1 inhibits CREB mediated activation of cyclic-AMP response element (CRE) containing promoters in a γ-secretase-dependent manner. We went on to find that the γ-cleaved N1 intracellular domain (N1ICD) sequesters nuclear CREB1α, inhibits cAMP/PKA-mediated neurite outgrowth and represses the expression of specific CREB regulated genes associated with learning and memory in primary cortical neurons. Similar transcriptional effects were observed with the N2ICD, N3ICD and N4ICDs. Together, these observations indicate that the effects of Notch on learning and memory are, at least in part, via an effect on CREB-regulated gene expression. PMID:25479589

  18. Targeting the annulus fibrosus of the intervertebral disc: Col1a2-Cre(ER)T mice show specific activity of Cre recombinase in the outer annulus fibrosus.

    PubMed

    Bedore, Jake; Quesnel, Katherine; Quinonez, Diana; Séguin, Cheryle A; Leask, Andrew

    2016-06-01

    Degeneration of the intervertebral disc (IVD) is a major underlying contributor to back pain-the single leading cause of disability worldwide. However, we possess a limited understanding of the etiology underlying IVD degeneration. To date, there are a limited number of mouse models that have been used to target proteins in specific compartments of the IVD to explore their functions in disc development, homeostasis and disease. Furthermore, the majority of reports exploring the composition and function of the outer encapsulating annulus fibrosus (AF) of the IVD have considered it as one tissue, without considering the numerous structural and functional differences existing between the inner and outer AF. In addition, no mouse models have yet been reported that enable specific targeting of genes within the outer AF. In the current report, we discuss these issues and demonstrate the localized activity of Cre recombinase in the IVD of Col1a2-Cre(ER)T;ROSA26mTmG mice possessing a tamoxifen-dependent Cre recombinase driven by a Cola2 promoter and distal enhancer and the mTmG fluorescent reporter. Following tamoxifen injection of 3-week-old Col1a2-Cre(ER)T;ROSA26mTmG mice, we show Cre activity specifically in the outer AF of the IVD, as indicated by expression of the GFP reporter. Thus, Col1a2-Cre(ER)T;ROSA26mTmG mice may prove to be a valuable tool in delineating the function of proteins in this unique compartment of the IVD, and in further exploring the compositional differences between the inner and outer AF in disc homeostasis, aging and disease. PMID:27173473

  19. Reliable Genetic Labeling of Adult-Born Dentate Granule Cells Using Ascl1CreERT2 and GlastCreERT2 Murine Lines

    PubMed Central

    Yang, Sung M.; Alvarez, Diego D.

    2015-01-01

    Newly generated dentate granule cells (GCs) are relevant for input discrimination in the adult hippocampus. Yet, their precise contribution to information processing remains unclear. To address this question, it is essential to develop approaches to precisely label entire cohorts of adult-born GCs. In this work, we used genetically modified mice to allow conditional expression of tdTomato (Tom) in adult-born GCs and characterized their development and functional integration. Ascl1CreERT2;CAGfloxStopTom and GlastCreERT2;CAGfloxStopTom mice resulted in indelible expression of Tom in adult neural stem cells and their lineage upon tamoxifen induction. Whole-cell recordings were performed to measure intrinsic excitability, firing behavior, and afferent excitatory connectivity. Developing GCs were also staged by the expression of early and late neuronal markers. The slow development of adult-born GCs characterized here is consistent with previous reports using retroviral approaches that have revealed that a mature phenotype is typically achieved after 6–8 weeks. Our findings demonstrate that Ascl1CreERT2 and GlastCreERT2 mouse lines enable simple and reliable labeling of adult-born GC lineages within restricted time windows. Therefore, these mice greatly facilitate tagging new neurons and manipulating their activity, required for understanding adult neurogenesis in the context of network remodeling, learning, and behavior. SIGNIFICANCE STATEMENT Our study shows that Ascl1CreERT2 and GlastCreERT2 mice lines can be used to label large cohorts of adult-born dentate granule cells with excellent time resolution. Neurons labeled in this manner display developmental and functional profiles that are in full agreement with previous findings using thymidine analogs and retroviral labeling, thus providing an alternative approach to tackle fundamental questions on circuit remodeling. Because of the massive neuronal targeting and the simplicity of this method, genetic labeling will

  20. Isolation of Novel CreERT2-Driver Lines in Zebrafish Using an Unbiased Gene Trap Approach

    PubMed Central

    Jungke, Peggy; Hammer, Juliane; Hans, Stefan; Brand, Michael

    2015-01-01

    Gene manipulation using the Cre/loxP-recombinase system has been successfully employed in zebrafish to study gene functions and lineage relationships. Recently, gene trapping approaches have been applied to produce large collections of transgenic fish expressing conditional alleles in various tissues. However, the limited number of available cell- and tissue-specific Cre/CreERT2-driver lines still constrains widespread application in this model organism. To enlarge the pool of existing CreERT2-driver lines, we performed a genome-wide gene trap screen using a Tol2-based mCherry-T2a-CreERT2 (mCT2aC) gene trap vector. This cassette consists of a splice acceptor and a mCherry-tagged variant of CreERT2 which enables simultaneous labeling of the trapping event, as well as CreERT2 expression from the endogenous promoter. Using this strategy, we generated 27 novel functional CreERT2-driver lines expressing in a cell- and tissue-specific manner during development and adulthood. This study summarizes the analysis of the generated CreERT2-driver lines with respect to functionality, expression, integration, as well as associated phenotypes. Our results significantly enlarge the existing pool of CreERT2-driver lines in zebrafish and combined with Cre–dependent effector lines, the new CreERT2-driver lines will be important tools to manipulate the zebrafish genome. PMID:26083735

  1. ELT and Consciousness-Raising

    ERIC Educational Resources Information Center

    Al-Jardani, Khalid Salim Saif

    2012-01-01

    The paper highlights the concept of consciousness-raising. It relates it to different aspects of ELT such as explicit teaching, language awareness, language acquisition and practice. How these terms are related to the concept of consciousness-raising within the English Language teaching. Its main aim is to help learners to notice for themselves…

  2. String Vacuous Overt Verb Raising.

    ERIC Educational Resources Information Center

    Koisumi, Masatoshi

    2000-01-01

    Presents evidence for overt verb raising in Japanese, thereby defending the validity of the head parameter with its two values (head initial and head final). Explores consequences of the overt verb raising analysis of Japanese to various aspects of syntactic theory. (Author/VWL)

  3. Oligodendroglial and Pan-neural Crest Expression of Cre Recombinase Directed by Sox10 Enhancer

    PubMed Central

    Stine, Zachary E.; Huynh, Jimmy L.; Loftus, Stacie K.; Gorkin, David U.; Salmasi, Amirali H.; Novak, Thomas; Purves, Todd; Miller, Ronald A.; Antonellis, Anthony; Gearhart, John P.; Pavan, William J.; McCallion, Andrew S.

    2010-01-01

    Utilizing a recently identified Sox10 distal enhancer directing Cre expression, we report S4F:Cre, a transgenic mouse line capable of inducing recombination in oligodendroglia and all examined neural crest derived tissues. Assayed using R26R:LacZ reporter mice expression was detected in neural crest derived tissues including the forming facial skeleton, dorsal root ganglia, sympathetic ganglia, enteric nervous system, aortae and melanoblasts, consistent with Sox10 expression. LacZ reporter expression was also detected in non-neural crest derived tissues including the oligodendrocytes and the ventral neural tube. This line provides appreciable differences in Cre expression pattern from other transgenic mouse lines that mark neural crest populations, including additional populations defined by the expression of other SoxE proteins. The S4F:Cre transgenic line will thus serve as a powerful tool for lineage tracing, gene function characterization and genome manipulation in these populations. PMID:19830815

  4. CDC Vital Signs: Making Health Care Safer -- Stop Infections from Lethal CRE Germs Now

    MedlinePlus

    ... 62 MB] Read the MMWR Science Clips Making Health Care Safer Stop Infections from Lethal CRE Germs Now ... to otherwise healthy people outside of medical facilities. Health Care Providers can Know if patients in your facility ...

  5. Monosynaptic circuit tracing in vivo through Cre-dependent targeting and complementation of modified rabies virus

    PubMed Central

    Wall, Nicholas R.; Wickersham, Ian R.; Cetin, Ali; De La Parra, Mauricio; Callaway, Edward M.

    2010-01-01

    We describe a powerful system for revealing the direct monosynaptic inputs to specific cell types in Cre-expressing transgenic mice through the use of Cre-dependent helper virus and a modified rabies virus. We generated helper viruses that target gene expression to Cre-expressing cells, allowing us to control initial rabies virus infection and subsequent monosynaptic retrograde spread. Investigators can use this system to elucidate the connections onto a desired cell type in a high-throughput manner, limited only by the availability of Cre mouse lines. This method allows for identification of circuits that would be extremely tedious or impossible to study with other methods and can be used to build subcircuit maps of inputs onto many different types of cells within the same brain region. Furthermore, by expressing various transgenes from the rabies genome, this system also has the potential to allow manipulation of targeted neuronal circuits without perturbing neighboring cells. PMID:21115815

  6. Raising Awareness in Science Education for Women (RAISE-W)

    NASA Astrophysics Data System (ADS)

    Faherty, Jacqueline K.; Holford, M.

    2014-01-01

    Raising Awareness in Science Education for Women (RAISE-W) is a 501c non profit corporation whose mission is to aid in increasing and retaining the number of women - especially underrepresented females - engaged in scientific teaching and research. Initiated by a Protein Chemist and an Astronomer, our ultimate goal has been to develop informational tools and create innovative outreach programs for women across all STEM fields. At present RAISE-W is recruiting women at the undergraduate, graduate, and early career stages to participate in a unique, 1-year, executive coaching program modeled after those used in the business sector.

  7. Multiple Levels of Affinity-Dependent DNA Discrimination in Cre-LoxP Recombination†

    PubMed Central

    Gelato, Kathy A.; Martin, Shelley S.; Wong, Scott; Baldwin, Enoch P.

    2010-01-01

    Cre recombinase residue Arg259 mediates a canonical bidentate hydrogen-bonded contact with Gua27 of its LoxP DNA substrate. Substituting Cyt8-Gua27 with the three other basepairs, to give LoxAT, LoxTA, and LoxGC, reduced Cre-mediated recombination in vitro, with the preference order of Gua27 >Ade27 ~ Thy27 ≫ Cyt27. While LoxAT and LoxTA exhibited 2.5-fold reduced affinity and 2.5-5-fold slower reaction rates, LoxGC was a barely functional substrate. Its maximum level of turnover was 6-fold reduced over other substrates, and it exhibited 8.5-fold reduced Cre binding and 6.3-fold slower turnover rate. With LoxP, the rate-limiting step for recombination occurs after protein-DNA complex assembly but before completion of the first strand exchange to form the Holliday junction (HJ) intermediate. With the mutant substrates, it occurs after HJ formation. Using an increased DNA-binding E262Q/E266Q “CreQQ” variant, all four substrates react more readily, but with much less difference between them, and maintained the earlier rate-limiting step. The data indicate that Cre discriminates substrates through differences in (i) concentration dependence of active complex assembly, (ii) turnover rate, and (iii) maximum yield of product at saturation, all of which are functions of the Cre-DNA binding interaction. CreQQ suppression of Lox mutant defects implies that coupling between binding and turnover involves a change in Cre subunit DNA affinities during the “conformational switch” that occurs prior to the second strand exchange. These results provide an example of how a DNA-binding enzyme can exert specificity via affinity modulation of conformational transitions that occur along its reaction pathway. PMID:17014075

  8. Multiple levels of affinity-dependent DNA discrimination in Cre-LoxP recombination.

    PubMed

    Gelato, Kathy A; Martin, Shelley S; Wong, Scott; Baldwin, Enoch P

    2006-10-10

    Cre recombinase residue Arg259 mediates a canonical bidentate hydrogen-bonded contact with Gua27 of its LoxP DNA substrate. Substituting Cyt8-Gua27 with the three other basepairs, to give LoxAT, LoxTA, and LoxGC, reduced Cre-mediated recombination in vitro, with the preference order of Gua27 > Ade27 approximately Thy27 > Cyt27. While LoxAT and LoxTA exhibited 2.5-fold reduced affinity and 2.5-5-fold slower reaction rates, LoxGC was a barely functional substrate. Its maximum level of turnover was 6-fold reduced over other substrates, and it exhibited 8.5-fold reduced Cre binding and 6.3-fold slower turnover rate. With LoxP, the rate-limiting step for recombination occurs after protein-DNA complex assembly but before completion of the first strand exchange to form the Holliday junction (HJ) intermediate. With the mutant substrates, it occurs after HJ formation. Using an increased DNA-binding E262Q/E266Q "CreQQ" variant, all four substrates react more readily, but with much less difference between them, and maintained the earlier rate-limiting step. The data indicate that Cre discriminates substrates through differences in (i) concentration dependence of active complex assembly, (ii) turnover rate, and (iii) maximum yield of product at saturation, all of which are functions of the Cre-DNA binding interaction. CreQQ suppression of Lox mutant defects implies that coupling between binding and turnover involves a change in Cre subunit DNA affinities during the "conformational switch" that occurs prior to the second strand exchange. These results provide an example of how a DNA-binding enzyme can exert specificity via affinity modulation of conformational transitions that occur along its reaction pathway. PMID:17014075

  9. Screening strategy to generate cell specific recombination: a case report with the RIP-Cre mice.

    PubMed

    Spinelli, Valeria; Martin, Céline; Dorchies, Emilie; Vallez, Emmanuelle; Dehondt, Hélène; Trabelsi, Mohamed-Sami; Tailleux, Anne; Caron, Sandrine; Staels, Bart

    2015-10-01

    Conditional gene knockout technology is a powerful tool to study the function of a gene in a specific tissue, organ or cell lineage. The most commonly used procedure applies the Cre-LoxP strategy, where the choice of the Cre driver promoter is critical to determine the efficiency and specificity of the system. However, a considered choice of an appropriate promoter does not always protect against the risk of unwanted recombination and the consequent deletion of the gene in other tissues than the desired one(s), due to phenomena of non-specific activation of the Cre transgene. Furthermore, the causes of these phenomena are not completely understood and this can potentially affect every strain of Cre-mice. In our study on the deletion of a same gene in two different tissues, we show that the incidence rate of non-specific recombination in unwanted tissues depends on the Cre driver strain, ranging from 100%, rendering it useless (aP2-Cre strain), to ~5%, which is still compatible with their use (RIP-Cre strain). The use of a simple PCR strategy conceived to detect this occurrence is indispensable when producing a tissue-specific knockout mouse. Therefore, when choosing the Cre-driver promoter, researchers not only have to be careful about its tissue-specificity and timing of activation, but should also include a systematical screening in order to exclude mice in which atypical recombination has occurred and to limit the unnecessary use of laboratory animals in uninterpretable experiments. PMID:26091792

  10. Strategic Plan for Higher Education, 2008-2017. Executive Summary

    ERIC Educational Resources Information Center

    Fingerhut, Eric D.

    2008-01-01

    If the State of Ohio is to grow and prosper, it must raise the education level of its population. The goal of this 10-year strategic plan is to raise the educational attainment of the state each year, and to close the gap between Ohio and competitor states and nations. To accomplish this goal stakeholders must do three things: (1) Graduate more…

  11. Strategic Plan for Higher Education, 2008-2017

    ERIC Educational Resources Information Center

    Fingerhut, Eric D.

    2008-01-01

    If the State of Ohio is to grow and prosper, it must raise the education level of its population. The goal of this 10-year strategic plan is to raise the educational attainment of the state each year, and to close the gap between Ohio and competitor states and nations. To accomplish this goal, stakeholders must do three things: (1) Graduate more…

  12. Evaluating Tissue-Specific Recombination in a Pdgfrα-CreERT2 Transgenic Mouse Line.

    PubMed

    O'Rourke, Megan; Cullen, Carlie L; Auderset, Loic; Pitman, Kimberley A; Achatz, Daniela; Gasperini, Robert; Young, Kaylene M

    2016-01-01

    In the central nervous system (CNS) platelet derived growth factor receptor alpha (PDGFRα) is expressed exclusively by oligodendrocyte progenitor cells (OPCs), making the Pdgfrα promoter an ideal tool for directing transgene expression in this cell type. Two Pdgfrα-CreERT2 mouse lines have been generated for this purpose which, when crossed with cre-sensitive reporter mice, allow the temporally restricted labelling of OPCs for lineage-tracing studies. These mice have also been used to achieve the deletion of CNS-specific genes from OPCs. However the ability of Pdgfrα-CreERT2 mice to induce cre-mediated recombination in PDGFRα+ cell populations located outside of the CNS has not been examined. Herein we quantify the proportion of PDGFRα+ cells that become YFP-labelled following Tamoxifen administration to adult Pdgfrα-CreERT2::Rosa26-YFP transgenic mice. We report that the vast majority (>90%) of PDGFRα+ OPCs in the CNS, and a significant proportion of PDGFRα+ stromal cells within the bone marrow (~38%) undergo recombination and become YFP-labelled. However, only a small proportion of the PDGFRα+ cell populations found in the sciatic nerve, adrenal gland, pituitary gland, heart, gastrocnemius muscle, kidney, lung, liver or intestine become YFP-labelled. These data suggest that Pdgfrα-CreERT2 transgenic mice can be used to achieve robust recombination in OPCs, while having a minimal effect on most PDGFRα+ cell populations outside of the CNS. PMID:27626928

  13. Spatially-Directed Assembly of a Heterotetrameric Cre-Lox Synapse Restricts Recombination Specificity

    PubMed Central

    Gelato, Kathy A.; Martin, Shelly S.; Liu, Patty H.; Saunders, April A.; Baldwin, Enoch P.

    2008-01-01

    SUMMARY The pseudo-fourfold homo-tetrameric synapse formed by Cre protein and target DNA restricts site-specific recombination to sequences containing dyad-symmetric Cre-binding repeats. Mixtures of engineered altered-specificity Cre monomers can form heterotetramers that recombine non-identical asymmetric sequences, allowing greater flexibility for target site selection in the genome of interest. However, the variety of tetramers allowed by random subunit association increases the chances of unintended reactivity at non-target sites. This problem can be circumvented by specifying a unique spatial arrangement of heterotetramer subunits. By reconfiguring inter-subunit protein-protein contacts, we directed the assembly of two different Cre monomers, each having a distinct DNA sequence specificity, in an alternating (ABAB) configuration. This designed heterotetramer preferentially recombined a particular pair of asymmetric Lox sites over other pairs, whereas a mixture of freely-associating subunits showed little bias. Alone, the engineered monomers had reduced reactivity towards both dyad-symmetric and asymmetric sites. Specificity arose because the organization of Cre-binding repeats of the preferred substrate matched the programmed arrangement of the subunits in the heterotetrameric synapse. Applying this “spatial matching” principle, Cre-mediated recombination can be directed to asymmetric DNA sequences with greater fidelity. PMID:18374357

  14. Split-Cre recombinase effectively monitors protein-protein interactions in living bacteria.

    PubMed

    O'Brien, Sean P; DeLisa, Matthew P

    2014-03-01

    The ability of Cre recombinase to excise genetic material has been used extensively for genome engineering in prokaryotic and eukaryotic cells. Recently, split-Cre fragments have been described that advance control of recombinase activity in mammalian cells. However, whether these fragments can be utilized for monitoring protein-protein interactions has not been reported. In this work, we developed a protein-fragment complementation assay (PCA) based on split-Cre for monitoring and engineering pairwise protein interactions in living Escherichia coli cells. This required creation of a dual-fluorescent reporter plasmid that permits visualization of reconstituted Cre recombinase activity by switching from red to green in the presence of an interacting protein pair. The resulting split-Cre PCA faithfully links cell fluorescence with differences in binding affinity, thereby allowing the facile isolation of high-affinity binders based on phenotype. Given the resolution of its activity and sensitivity to interactions, our system may prove a viable option for poorly expressed or weakly interacting protein pairs that evade detection in other PCA formats. Based on these findings, we anticipate that our split-Cre PCA will become a highly complementary and useful new addition to the protein-protein interaction toolbox. PMID:24390935

  15. Monodispersity of recombinant Cre recombinase correlates with its effectiveness in vivo

    PubMed Central

    Capasso, Paola; Aliprandi, Marisa; Ossolengo, Giuseppe; Edenhofer, Frank; de Marco, Ario

    2009-01-01

    Background Cre recombinase is a common reagent used for the in vivo on/off switching of the expression of target genes flanked by loxP sites. In particular, recombinant TAT-Cre fusion constructs purified from bacteria have been used to promote the cell uptake of the enzyme. However, the recovery of active TAT-Cre remains a demanding process and its specific activity varies significantly among batches, making difficult data comparison. Results We noticed a strong correlation between recombinase activity and enzyme monodispersity. The existence of such correlation enabled us to indirectly monitor the TAT-Cre recombinase activity during the multi-step purification process by measuring its monodispersity, a parameter detectable by means of a spectrofluorimetric assay that allows the calculation of the Aggregation Index (AI) in an easy and rapid way. AI values were recorded after each purification passage to identify the critical steps and to choose optimal alternatives for chromatographic conditions, desalting procedures, and protocols for bacterial endotoxin removal. Furthermore, the effect of metal ions and temperature on TAT-Cre aggregation and inactivation was characterized in vitro. Finally, we optimized the enzyme delivery protocol in vivo by following the accumulation tuning of the reporter protein β-catenin. Conclusion A rational purification protocol for TAT-Cre has been developed by choosing the options that minimize the enzyme aggregation. Our data suggest that AI measurement should support the optimization of any protocol aiming at the recovery of monodispersed protein. PMID:19747375

  16. Spatially directed assembly of a heterotetrameric Cre-Lox synapse restricts recombination specificity.

    PubMed

    Gelato, Kathy A; Martin, Shelley S; Liu, Patty H; Saunders, April A; Baldwin, Enoch P

    2008-05-01

    The pseudo-fourfold homotetrameric synapse formed by Cre protein and target DNA restricts site-specific recombination to sequences containing dyad-symmetric Cre-binding repeats. Mixtures of engineered altered-specificity Cre monomers can form heterotetramers that recombine nonidentical asymmetric sequences, allowing greater flexibility for target site selection in the genome of interest. However, the variety of tetramers allowed by random subunit association increases the chances of unintended reactivity at nontarget sites. This problem can be circumvented by specifying a unique spatial arrangement of heterotetramer subunits. By reconfiguring intersubunit protein-protein contacts, we directed the assembly of two different Cre monomers, each having a distinct DNA sequence specificity, in an alternating (ABAB) configuration. This designed heterotetramer preferentially recombined a particular pair of asymmetric Lox sites over other pairs, whereas a mixture of freely associating subunits showed little bias. Alone, the engineered monomers had reduced reactivity towards both dyad-symmetric and asymmetric sites. Specificity arose because the organization of Cre-binding repeats of the preferred substrate matched the programmed arrangement of the subunits in the heterotetrameric synapse. When this "spatial matching" principle is applied, Cre-mediated recombination can be directed to asymmetric DNA sequences with greater fidelity. PMID:18374357

  17. Construction of Marker-Free Transgenic Strains of Chlamydomonas reinhardtii Using a Cre/loxP-Mediated Recombinase System

    PubMed Central

    Kasai, Yuki; Harayama, Shigeaki

    2016-01-01

    The Escherichia coli bacteriophage P1 encodes a site-specific recombinase called Cre and two 34-bp target sites of Cre recombinase called loxP. The Cre/loxP system has been used to achieve targeted insertion and precise deletion in many animal and plant genomes. The Cre/loxP system has particularly been used for the removal of selectable marker genes to create marker-free transgenic organisms. For the first time, we applied the Cre/loxP-mediated site-specific recombination system to Chlamydomonas reinhardtii to construct marker-free transgenic strains. Specifically, C. reinhardtii strains cc4350 and cc124 carrying an aphVIII expression cassette flanked by two direct repeats of loxP were constructed. Separately, a synthetic Cre recombinase gene (CrCRE), the codons of which were optimized for expression in C. reinhardtii, was synthesized, and a CrCRE expression cassette was introduced into strain cc4350 carrying a single copy of the loxP-flanked aphVIII expression cassette. Among 46 transformants carrying the CrCRE expression cassette stably, the excision of aphVIII by CrCre recombinase was observed only in one transformant. We then constructed an expression cassette of an in-frame fusion of ble to CrCRE via a short linker peptide. The product of ble (Ble) is a bleomycin-binding protein that confers resistance to bleomycin-related antibiotics such as Zeocin and localizes in the nucleus. Therefore, the ble-(linker)-CrCRE fusion protein is expected to localize in the nucleus. When the ble-(linker)-CrCRE expression cassette was integrated into the genome of strain cc4350 carrying a single copy of the loxP-flanked aphVIII expression cassette, CrCre recombinase-mediated excision of the aphVIII expression cassette was observed at a frequency higher than that in stable transformants of the CrCRE expression cassette. Similarly, from strain cc124 carrying a single loxP-flanked aphVIII expression cassette, the aphVIII expression cassette was successfully excised after

  18. Construction of Marker-Free Transgenic Strains of Chlamydomonas reinhardtii Using a Cre/loxP-Mediated Recombinase System.

    PubMed

    Kasai, Yuki; Harayama, Shigeaki

    2016-01-01

    The Escherichia coli bacteriophage P1 encodes a site-specific recombinase called Cre and two 34-bp target sites of Cre recombinase called loxP. The Cre/loxP system has been used to achieve targeted insertion and precise deletion in many animal and plant genomes. The Cre/loxP system has particularly been used for the removal of selectable marker genes to create marker-free transgenic organisms. For the first time, we applied the Cre/loxP-mediated site-specific recombination system to Chlamydomonas reinhardtii to construct marker-free transgenic strains. Specifically, C. reinhardtii strains cc4350 and cc124 carrying an aphVIII expression cassette flanked by two direct repeats of loxP were constructed. Separately, a synthetic Cre recombinase gene (CrCRE), the codons of which were optimized for expression in C. reinhardtii, was synthesized, and a CrCRE expression cassette was introduced into strain cc4350 carrying a single copy of the loxP-flanked aphVIII expression cassette. Among 46 transformants carrying the CrCRE expression cassette stably, the excision of aphVIII by CrCre recombinase was observed only in one transformant. We then constructed an expression cassette of an in-frame fusion of ble to CrCRE via a short linker peptide. The product of ble (Ble) is a bleomycin-binding protein that confers resistance to bleomycin-related antibiotics such as Zeocin and localizes in the nucleus. Therefore, the ble-(linker)-CrCRE fusion protein is expected to localize in the nucleus. When the ble-(linker)-CrCRE expression cassette was integrated into the genome of strain cc4350 carrying a single copy of the loxP-flanked aphVIII expression cassette, CrCre recombinase-mediated excision of the aphVIII expression cassette was observed at a frequency higher than that in stable transformants of the CrCRE expression cassette. Similarly, from strain cc124 carrying a single loxP-flanked aphVIII expression cassette, the aphVIII expression cassette was successfully excised after

  19. Distracted Driving Raises Crash Risk

    MedlinePlus

    ... Raises Crash Risk Video technology and in-vehicle sensors showed that distracted driving, especially among new drivers, ... whenever the cars were moving. A suite of sensors recorded acceleration, sudden braking or swerving, and other ...

  20. Expression and regulation of CReMM, a chromodomain helicase-DNA-binding (CHD), in marrow stroma derived osteoprogenitors.

    PubMed

    Marom, R; Shur, I; Hager, G L; Benayahu, D

    2006-06-01

    This study follows the expression of CReMM, a new CHD family member, in osteoprogenitors. CReMM expression was analyzed in primary cultured mesnchymal cells from rat and human. Analysis in ex vivo cultured marrow stromal cells (MSC) from rats revealed higher level of CReMM in cells from young (3 months), when compared to cells from old (15 months) rats. CReMM level was higher in human MSC then in mature trabecular bone cells (TBC). Within the MSC population, osteogenic clones showed higher levels of CReMM then non-osteogenic ones. We used bone marrow derived osteogenic cell line (MBA-15) to elaborate on the regulation of CReMM expression in correlation with cell proliferation and co-expression with alkaline phosphatase (ALK). CReMM is highly expressed in proliferating cells and is inversely related to expression of ALK. MBA-15 cells were challenged with dexamethasone (Dex) or 17beta-estradiol and quantification of CReMM at the protein (ELISA) and mRNA (RT-PCR) levels had shown that Dex upregulated CReMM levels. Since CReMM is regulated by Dex, we analyzed the interaction of CReMM with the glucocorticoid receptor (GR), which mediates Dex action. Co-immunopercipitation (Co-IP) demonstrated an association between CReMM and GR. In summary, CReMM is a CHD protein expressed by osteoprogenitors, and we suggest it plays a role in mediating transcriptional response to hormones that coordinate osteoblast function. PMID:16523501

  1. In Vivo Performance and Properties of Tamoxifen Metabolites for CreERT2 Control

    PubMed Central

    Dolbois, Aymeric; Blazkova, Kristyna; Hess, Christopher; Low, Larry W. L.; Burger, Sibylle; Samson, Natasha; Carney, Tom J.; Bartunek, Petr; Nevado, Cristina

    2016-01-01

    Mutant Estrogen Receptor (ERT2) ligand-binding domain fusions with Cre recombinase are a key tool for spatio-temporally controlled genetic recombination with the Cre/lox system. CreERT2 is efficiently activated in a concentration-dependent manner by the Tamoxifen metabolite trans-4-OH-Tamoxifen (trans-4-OHT). Reproducible and efficient Cre/lox experimentation is hindered by the gradual loss of CreERT2 induction potency upon prolonged storage of dissolved trans-4-OHT, which potentially results from gradual trans-to-cis isomerization or degradation. Here, we combined zebrafish CreERT2 recombination experiments and cell culture assays to document the gradual activity loss of trans-4-OHT and describe the alternative Tamoxifen metabolite Endoxifen as more stable alternative compound. Endoxifen retains potent activation upon prolonged storage (3 months), yet consistently induces half the ERT2 domain fusion activity compared to fresh trans-4-OHT. Using 1H-NMR analysis, we reveal that trans-4-OHT isomerization is undetectable upon prolonged storage in either DMSO or Ethanol, ruling out isomer transformation as cause for the gradual loss of trans-4-OHT activity. We further establish that both trans-4-OHT and Endoxifen are insensitive to light exposure under regular laboratory handling conditions. We attribute the gradual loss of trans-4-OHT potency to precipitation over time, and show that heating of aged trans-4-OHT aliquots reinstates their CreERT2 induction potential. Our data establish Endoxifen as potent and reproducible complementary compound to 4-OHT to control ERT2 domain fusion proteins in vivo, and provide a framework for efficient chemically controlled recombination experiments. PMID:27077909

  2. In Vivo Performance and Properties of Tamoxifen Metabolites for CreERT2 Control.

    PubMed

    Felker, Anastasia; Nieuwenhuize, Susan; Dolbois, Aymeric; Blazkova, Kristyna; Hess, Christopher; Low, Larry W L; Burger, Sibylle; Samson, Natasha; Carney, Tom J; Bartunek, Petr; Nevado, Cristina; Mosimann, Christian

    2016-01-01

    Mutant Estrogen Receptor (ERT2) ligand-binding domain fusions with Cre recombinase are a key tool for spatio-temporally controlled genetic recombination with the Cre/lox system. CreERT2 is efficiently activated in a concentration-dependent manner by the Tamoxifen metabolite trans-4-OH-Tamoxifen (trans-4-OHT). Reproducible and efficient Cre/lox experimentation is hindered by the gradual loss of CreERT2 induction potency upon prolonged storage of dissolved trans-4-OHT, which potentially results from gradual trans-to-cis isomerization or degradation. Here, we combined zebrafish CreERT2 recombination experiments and cell culture assays to document the gradual activity loss of trans-4-OHT and describe the alternative Tamoxifen metabolite Endoxifen as more stable alternative compound. Endoxifen retains potent activation upon prolonged storage (3 months), yet consistently induces half the ERT2 domain fusion activity compared to fresh trans-4-OHT. Using 1H-NMR analysis, we reveal that trans-4-OHT isomerization is undetectable upon prolonged storage in either DMSO or Ethanol, ruling out isomer transformation as cause for the gradual loss of trans-4-OHT activity. We further establish that both trans-4-OHT and Endoxifen are insensitive to light exposure under regular laboratory handling conditions. We attribute the gradual loss of trans-4-OHT potency to precipitation over time, and show that heating of aged trans-4-OHT aliquots reinstates their CreERT2 induction potential. Our data establish Endoxifen as potent and reproducible complementary compound to 4-OHT to control ERT2 domain fusion proteins in vivo, and provide a framework for efficient chemically controlled recombination experiments. PMID:27077909

  3. Heme oxygenase-1 expression protects the heart from acute injury caused by inducible Cre recombinase

    PubMed Central

    Hull, Travis D.; Bolisetty, Subashini; DeAlmeida, Angela; Litovsky, Silvio H.; Prabhu, Sumanth D.; Agarwal, Anupam; George, James F.

    2013-01-01

    The protective effect of heme oxygenase-1 (HO-1) expression in cardiovascular disease has been previously demonstrated using transgenic animal models in which HO-1 is constitutively overexpressed in the heart. However, the temporal requirements for protection by HO-1 induction relative to injury have not been investigated, but are essential to employ HO-1 as a therapeutic strategy in human cardiovascular disease states. Therefore, we generated mice with cardiac-specific, tamoxifen (TAM)-inducible overexpression of a human HO-1 (hHO-1) transgene (MHC-HO-1 mice) by breeding mice with cardiac-specific expression of a TAM-inducible Cre recombinase (MHC-Cre mice) with mice containing an hHO-1 transgene preceded by a floxed stop signal (CBA-flox mice). MHC-HO-1 overexpress the HO-1 gene and enzymatically protein following TAM administration (40 mg/kg body weight on two consecutive days). In MHC-Cre controls, TAM administration leads to severe, acute cardiac toxicity, cardiomyocyte necrosis, and 80% mortality by day 3. This cardiac toxicity is accompanied by a significant increase in inflammatory cells in the heart that are predominantly neutrophils. In MHC-HO-1 mice, HO-1 overexpression ameliorates the depression of cardiac function and high mortality rate observed in MHC-Cre mice following TAM administration and attenuates cardiomyocyte necrosis and neutrophil infiltration. These results highlight that HO-1 induction is sufficient to prevent the depression of cardiac function observed in mice with TAM-inducible Cre recombinase expression by protecting the heart from necrosis and neutrophil infiltration. These findings are important because MHC-Cre mice are widely used in cardiovascular research despite the limitations imposed by Cre-induced cardiac toxicity and also because inflammation is an important pathological component of many human cardiovascular diseases. PMID:23732814

  4. Heme oxygenase-1 expression protects the heart from acute injury caused by inducible Cre recombinase.

    PubMed

    Hull, Travis D; Bolisetty, Subhashini; DeAlmeida, Angela C; Litovsky, Silvio H; Prabhu, Sumanth D; Agarwal, Anupam; George, James F

    2013-08-01

    The protective effect of heme oxygenase-1 (HO-1) expression in cardiovascular disease has been previously demonstrated using transgenic animal models in which HO-1 is constitutively overexpressed in the heart. However, the temporal requirements for protection by HO-1 induction relative to injury have not been investigated, but are essential to employ HO-1 as a therapeutic strategy in human cardiovascular disease states. Therefore, we generated mice with cardiac-specific, tamoxifen (TAM)-inducible overexpression of a human HO-1 (hHO-1) transgene (myosin heavy chain (MHC)-HO-1 mice) by breeding mice with cardiac-specific expression of a TAM-inducible Cre recombinase (MHC-Cre mice), with mice containing an hHO-1 transgene preceded by a floxed-stop signal. MHC-HO-1 mice overexpress HO-1 mRNA and the enzymatically active protein following TAM administration (40 mg/kg body weight on 2 consecutive days). In MHC-Cre controls, TAM administration leads to severe, acute cardiac toxicity, cardiomyocyte necrosis, and 80% mortality by day 3. This cardiac toxicity is accompanied by a significant increase in inflammatory cells in the heart that are predominantly neutrophils. In MHC-HO-1 mice, HO-1 overexpression ameliorates the depression of cardiac function and high mortality rate observed in MHC-Cre mice following TAM administration and attenuates cardiomyocyte necrosis and neutrophil infiltration. These results highlight that HO-1 induction is sufficient to prevent the depression of cardiac function observed in mice with TAM-inducible Cre recombinase expression by protecting the heart from necrosis and neutrophil infiltration. These findings are important because MHC-Cre mice are widely used in cardiovascular research despite the limitations imposed by Cre-induced cardiac toxicity, and also because inflammation is an important pathological component of many human cardiovascular diseases. PMID:23732814

  5. Quantitative comparison of cis-regulatory element (CRE) activities in transgenic Drosophila melanogaster.

    PubMed

    Rogers, William A; Williams, Thomas M

    2011-01-01

    Gene expression patterns are specified by cis-regulatory element (CRE) sequences, which are also called enhancers or cis-regulatory modules. A typical CRE possesses an arrangement of binding sites for several transcription factor proteins that confer a regulatory logic specifying when, where, and at what level the regulated gene(s) is expressed. The full set of CREs within an animal genome encodes the organism's program for development, and empirical as well as theoretical studies indicate that mutations in CREs played a prominent role in morphological evolution. Moreover, human genome wide association studies indicate that genetic variation in CREs contribute substantially to phenotypic variation. Thus, understanding regulatory logic and how mutations affect such logic is a central goal of genetics. Reporter transgenes provide a powerful method to study the in vivo function of CREs. Here a known or suspected CRE sequence is coupled to heterologous promoter and coding sequences for a reporter gene encoding an easily observable protein product. When a reporter transgene is inserted into a host organism, the CRE's activity becomes visible in the form of the encoded reporter protein. P-element mediated transgenesis in the fruit fly species Drosophila (D.) melanogaster has been used for decades to introduce reporter transgenes into this model organism, though the genomic placement of transgenes is random. Hence, reporter gene activity is strongly influenced by the local chromatin and gene environment, limiting CRE comparisons to being qualitative. In recent years, the phiC31 based integration system was adapted for use in D. melanogaster to insert transgenes into specific genome landing sites. This capability has made the quantitative measurement of gene and, relevant here, CRE activity feasible. The production of transgenic fruit flies can be outsourced, including phiC31-based integration, eliminating the need to purchase expensive equipment and/or have proficiency at

  6. Raising HDL cholesterol in women

    PubMed Central

    Eapen, Danny J; Kalra, Girish L; Rifai, Luay; Eapen, Christina A; Merchant, Nadya; Khan, Bobby V

    2010-01-01

    High-density lipoprotein cholesterol (HDL-C) concentration is essential in the determination of coronary heart disease (CHD) risk in women. This is especially true in the postmenopausal state, where lipid profiles and CHD risk mimic that of age-matched men. Thus, interventions designed to reduce CHD risk by raising HDL-C levels may have particular significance during the transition to menopause. This review discusses HDL-C-raising therapies and the role of HDL in the primary prevention of CHD in women. Lifestyle-based interventions such as dietary change, aerobic exercise regimens, and smoking cessation are initial steps that are effective in raising HDL-C, and available data suggest women respond similarly to men with these interventions. When combined with pharmacotherapy, the effects of these lifestyle alterations are further amplified. Though studies demonstrating gender-specific differences in therapy are limited, niacin continues to be the most effective agent in raising HDL-C levels, especially when used in combination with fibrate or statin therapy. Emerging treatments such as HDL mimetic therapy show much promise in further raising HDL-C levels and improving cardiovascular outcomes. PMID:21072287

  7. Cell-Specific Cre Strains For Genetic Manipulation in Salivary Glands

    PubMed Central

    Maruyama, Eri O.; Aure, Marit H.; Xie, Xiaoling; Myal, Yvonne; Gan, Lin; Ovitt, Catherine E.

    2016-01-01

    The secretory acinar cells of the salivary gland are essential for saliva secretion, but are also the cell type preferentially lost following radiation treatment for head and neck cancer. The source of replacement acinar cells is currently a matter of debate. There is evidence for the presence of adult stem cells located within specific ductal regions of the salivary glands, but our laboratory recently demonstrated that differentiated acinar cells are maintained without significant stem cell contribution. To enable further investigation of salivary gland cell lineages and their origins, we generated three cell-specific Cre driver mouse strains. For genetic manipulation in acinar cells, an inducible Cre recombinase (Cre-ER) was targeted to the prolactin-induced protein (Pip) gene locus. Targeting of the Dcpp1 gene, encoding demilune cell and parotid protein, labels intercalated duct cells, a putative site of salivary gland stem cells, and serous demilune cells of the sublingual gland. Duct cell-specific Cre expression was attempted by targeting the inducible Cre to the Tcfcp2l1 gene locus. Using the R26Tomato Red reporter mouse, we demonstrate that these strains direct inducible, cell-specific expression. Genetic tracing of acinar cells using PipGCE supports the recent finding that differentiated acinar cells clonally expand. Moreover, tracing of intercalated duct cells expressing DcppGCE confirms evidence of duct cell proliferation, but further analysis is required to establish that renewal of secretory acinar cells is dependent on stem cells within these ducts. PMID:26751783

  8. Deletion of creB in Aspergillus oryzae increases secreted hydrolytic enzyme activity.

    PubMed

    Hunter, A J; Morris, T A; Jin, B; Saint, C P; Kelly, J M

    2013-09-01

    Aspergillus oryzae has been used in the food and beverage industry for centuries, and industrial strains have been produced by multiple rounds of selection. Targeted gene deletion technology is particularly useful for strain improvement in such strains, particularly when they do not have a well-characterized meiotic cycle. Phenotypes of an Aspergillus nidulans strain null for the CreB deubiquitinating enzyme include effects on growth and repression, including increased activity levels of various enzymes. We show that Aspergillus oryzae contains a functional homologue of the CreB deubiquitinating enzyme and that a null strain shows increased activity levels of industrially important secreted enzymes, including cellulases, xylanases, amylases, and proteases, as well as alleviated inhibition of spore germination on glucose medium. Reverse transcription-quantitative PCR (RT-qPCR) analysis showed that the increased levels of enzyme activity in both Aspergillus nidulans and Aspergillus oryzae are mirrored at the transcript level, indicating transcriptional regulation. We report that Aspergillus oryzae DAR3699, originally isolated from soy fermentation, has a similar phenotype to that of a creB deletion mutant of the RIB40 strain, and it contains a mutation in the creB gene. Collectively, the results for Aspergillus oryzae, Aspergillus nidulans, Trichoderma reesei, and Penicillium decumbens show that deletion of creB may be broadly useful in diverse fungi for increasing production of a variety of enzymes. PMID:23835170

  9. Postnatal lineage mapping of follicular melanocytes with the Tyr::CreER(T) (2) transgene.

    PubMed

    Harris, Melissa L; Pavan, William J

    2013-03-01

    One of the main advantages of using inducible and conditional transgenes to study pigment cell biology is that they allow for genetic manipulation within melanocytes after roles in general neural crest or melanoblast development have been fulfilled. Specifically, we focus here on the ability of the Tyr::CreER(T) (2) transgenic line to alter genes within follicular melanocytes postnatally. Using the Gt(ROSA)26Sor(tm1sor) reporter allele, we present in detail the expression and activity of Tyr::CreER(T) (2) when induced during hair morphogenesis and adult hair cycling. We find that despite similarities in expression pattern to endogenous TYR, Tyr::CreER(T) (2) is effective at targeting both undifferentiated and differentiated melanocytes within the hair follicle. We also find that Tyr::CreER(T) (2) provides the highest levels of recombination when induced during the early phases of hair growth. In conclusion, the descriptions provided here will guide future analyses of gene function within the melanocyte system of the hair follicle when using this Tyr::CreER(T) (2) transgene. PMID:23176440

  10. Molecular switch of Cre/loxP for radiation modulated gene therapy on hepatoma

    NASA Astrophysics Data System (ADS)

    Hsieh, Ya-Ju; Chen, Fu-Du; Wang, Fu Hui; Ke, Chien Chih; Wang, Hsin-Ell; Liu, Ren-Shyan

    2007-02-01

    For the purpose of enhancement of AFP promoter for the use of radiation modulated gene therapy for hepatocellular carcinoma (HCC), we combined hepatitis B virus (HBV) enhancer II with AFP promoter which shows the selectivity to the target cells to control the Cre/loxP system. Different gene constructs, pE4luc, pE4Tk, EIIAPA-Cre, E4CMV-STOP-Tk and chimeric promoters combined with HBV enhancer were constructed and transfected into HepG2, HeLa and NIH-3T3 cell lines. Cell experiments revealed that E4 enhancer responses to radiation best after 60 h irradiation at a dose range of 5-7 Gy in HepG2 stable clone. The EIIAPA promoter provided high specificity to hepatoma and activated the Cre downstream and removed the stop cassette only in hepatoma cells. After removal of the stop cassette, the E4 response to radiation could encode more Tk protein and kill more tumor cells. In summary, the chimeric EIIAPA promoter can stringently control the expression of Cre recombinase only in HCC. The radiation effect of the EIIAPA-Cre and E4CMV-STOP-Tk system shows promising results in terms of cell survival of HCC.

  11. Neuropilin-2 genomic elements drive cre recombinase expression in primitive blood, vascular and neuronal lineages.

    PubMed

    Wiszniak, Sophie; Scherer, Michaela; Ramshaw, Hayley; Schwarz, Quenten

    2015-11-01

    We have established a novel Cre mouse line, using genomic elements encompassing the Nrp2 locus, present within a bacterial artificial chromosome clone. By crossing this Cre driver line to R26R LacZ reporter mice, we have documented the temporal expression and lineage traced tissues in which Cre is expressed. Nrp2-Cre drives expression in primitive blood cells arising from the yolk sac, venous and lymphatic endothelial cells, peripheral sensory ganglia, and the lung bud. This mouse line will provide a new tool to researchers wishing to study the development of various tissues and organs in which this Cre driver is expressed, as well as allow tissue-specific knockout of genes of interest to study protein function. This work also presents the first evidence for expression of Nrp2 protein in a mesodermal progenitor with restricted hematopoietic potential, which will significantly advance the study of primitive erythropoiesis. genesis 53:709-717, 2015. © 2015 Wiley Periodicals, Inc. PMID:26454009

  12. Deletion of creB in Aspergillus oryzae Increases Secreted Hydrolytic Enzyme Activity

    PubMed Central

    Hunter, A. J.; Morris, T. A.; Jin, B.; Saint, C. P.

    2013-01-01

    Aspergillus oryzae has been used in the food and beverage industry for centuries, and industrial strains have been produced by multiple rounds of selection. Targeted gene deletion technology is particularly useful for strain improvement in such strains, particularly when they do not have a well-characterized meiotic cycle. Phenotypes of an Aspergillus nidulans strain null for the CreB deubiquitinating enzyme include effects on growth and repression, including increased activity levels of various enzymes. We show that Aspergillus oryzae contains a functional homologue of the CreB deubiquitinating enzyme and that a null strain shows increased activity levels of industrially important secreted enzymes, including cellulases, xylanases, amylases, and proteases, as well as alleviated inhibition of spore germination on glucose medium. Reverse transcription-quantitative PCR (RT-qPCR) analysis showed that the increased levels of enzyme activity in both Aspergillus nidulans and Aspergillus oryzae are mirrored at the transcript level, indicating transcriptional regulation. We report that Aspergillus oryzae DAR3699, originally isolated from soy fermentation, has a similar phenotype to that of a creB deletion mutant of the RIB40 strain, and it contains a mutation in the creB gene. Collectively, the results for Aspergillus oryzae, Aspergillus nidulans, Trichoderma reesei, and Penicillium decumbens show that deletion of creB may be broadly useful in diverse fungi for increasing production of a variety of enzymes. PMID:23835170

  13. NASA Strategic Plan

    NASA Technical Reports Server (NTRS)

    1996-01-01

    The aforementioned strategic decisions and the overarching direction for America's aeronautics and space program are addressed in the Strategic Plan. Our Strategic Plan is critical to our ability to meet the challenges of this new era and deliver a vibrant aeronautics and space program that strengthens and inspires the Nation. The Plan is our top-level strategy.

  14. Strategic Leadership Reconsidered

    ERIC Educational Resources Information Center

    Davies, Brent; Davies, Barbara J.

    2005-01-01

    This paper will address the challenge of how strategic leadership can be defined and articulated to provide a framework for developing a strategically focused school drawing on a NCSL research project. The paper is structured into three main parts. Part one outlines the elements that comprise a strategically focused school, develops an…

  15. Establishment of a tamoxifen-inducible Cre-driver mouse strain for widespread and temporal genetic modification in adult mice.

    PubMed

    Ichise, Hirotake; Hori, Akiko; Shiozawa, Seiji; Kondo, Saki; Kanegae, Yumi; Saito, Izumu; Ichise, Taeko; Yoshida, Nobuaki

    2016-07-29

    Temporal genetic modification of mice using the ligand-inducible Cre/loxP system is an important technique that allows the bypass of embryonic lethal phenotypes and access to adult phenotypes. In this study, we generated a tamoxifen-inducible Cre-driver mouse strain for the purpose of widespread and temporal Cre recombination. The new line, named CM32, expresses the GFPneo-fusion gene in a wide variety of tissues before FLP recombination and tamoxifen-inducible Cre after FLP recombination. Using FLP-recombined CM32 mice (CM32Δ mice) and Cre reporter mouse lines, we evaluated the efficiency of Cre recombination with and without tamoxifen administration to adult mice, and found tamoxifen-dependent induction of Cre recombination in a variety of adult tissues. In addition, we demonstrated that conditional activation of an oncogene could be achieved in adults using CM32Δ mice. CM32Δ;T26 mice, which harbored a Cre recombination-driven, SV40 large T antigen-expressing transgene, were viable and fertile. No overt phenotype was found in the mice up to 3 months after birth. Although they displayed pineoblastomas (pinealoblastomas) and/or thymic enlargement due to background Cre recombination by 6 months after birth, they developed epidermal hyperplasia when administered tamoxifen. Collectively, our results suggest that the CM32Δ transgenic mouse line can be applied to the assessment of adult phenotypes in mice with loxP-flanked transgenes. PMID:26923756

  16. Site-specific DNA excision in transgenic rice with a cell-permeable cre recombinase.

    PubMed

    Cao, Ming-Xia; Huang, Jian-Qiu; Yao, Quan-Hong; Liu, Sheng-Jun; Wang, Cheng-Long; Wei, Zhi-Ming

    2006-01-01

    The removal of selected marker genes from transgenic plants is necessary to address biosafety concerns and to carry out further experiments with transgenic organisms. In the present study, the 12-amino-acid membrane translocation sequence (MTS) from the Kaposi fibroblast growth factor (FGF)-4 was used as a carrier to deliver enzymatically active Cre proteins into living plant cells, and to produce a site-specific DNA excision in transgenic rice plants. The process, which made cells permeable to Cre recombinase-mediated DNA recombination, circumvented the need to express Cre under spatiotemporal control and was proved to be a simple and efficient system to achieve marker-free transgenic plants. The ultimate aim of the present study is to develop commercial rice cultivars free from selected marker genes to hasten public acceptance of transgenic crops. PMID:16382182

  17. Cre-driven optogenetics in the heterogeneous genetic panorama of the VTA.

    PubMed

    Pupe, Stéfano; Wallén-Mackenzie, Åsa

    2015-06-01

    The selectivity of optogenetics commonly relies on genetic promoters to manipulate specific populations of neurons through the use of Cre-driver lines. All studies performed in the ventral tegmental area (VTA) so far have utilized promoters present in groups of cells that release dopamine (DA), GABA, or glutamate. However, neurons that co-release neurotransmitters and variabilities within groups of neurons that release the same neurotransmitter present challenges when evaluating the results. Further complexity is introduced by ectopic expression patterns often occurring in transgenic Cre-drivers. New perspectives could be unfolded by identifying and selecting different types of promoter for driving the Cre recombinase. Here, we discuss some promising candidates and highlight the advantages or disadvantages of different methods for creating novel transgenic lines. PMID:25962754

  18. Evaluation of Remel Spectra CRE Agar for Detection of Carbapenem-Resistant Bacteria from Rectal Swabs Obtained from Residents of a Long-Term-Care Facility

    PubMed Central

    Urban, Carl M.; Kreiswirth, Barry N.; Chen, Liang; Osorio, Giuliana; Kopacz, Joanna; Labaze, Georges; Segal-Maurer, Sorana

    2015-01-01

    We compared the Remel Spectra CRE agar plate to CDC standard methodology for the isolation of carbapenem-resistant Enterobacteriaceae (CRE) from 300 rectal swab specimens obtained from patients residing in a long-term-care facility (LTCF). Multiplex PCR experiments were performed on isolates to identify specific Klebsiella pneumoniae carbapenemases (KPC) and additional β-lactamases. Of the 300 patients, 72 (24%) harbored CRE and were PCR positive for KPC enzymes. The Remel Spectra CRE plates detected KPC-type CRE in isolates from 70 of 72 patients (97.2%), while the CDC method detected CRE in 56 of 72 (77.8%). CRE identification results were available in 18 h compared to 36 h for the CDC method. Remel Spectra CRE agar plates can provide useful means for a fast and reliable method for detecting KPC-type CRE and for accelerated institution of appropriate infection control precautions. PMID:26085613

  19. Overlapping ETS and CRE Motifs ((G/C)CGGAAGTGACGTCA) preferentially bound by GABPα and CREB proteins.

    PubMed

    Chatterjee, Raghunath; Zhao, Jianfei; He, Ximiao; Shlyakhtenko, Andrey; Mann, Ishminder; Waterfall, Joshua J; Meltzer, Paul; Sathyanarayana, B K; FitzGerald, Peter C; Vinson, Charles

    2012-10-01

    Previously, we identified 8-bps long DNA sequences (8-mers) that localize in human proximal promoters and grouped them into known transcription factor binding sites (TFBS). We now examine split 8-mers consisting of two 4-mers separated by 1-bp to 30-bps (X(4)-N(1-30)-X(4)) to identify pairs of TFBS that localize in proximal promoters at a precise distance. These include two overlapping TFBS: the ETS⇔ETS motif ((C/G)CCGGAAGCGGAA) and the ETS⇔CRE motif ((C/G)CGGAAGTGACGTCAC). The nucleotides in bold are part of both TFBS. Molecular modeling shows that the ETS⇔CRE motif can be bound simultaneously by both the ETS and the B-ZIP domains without protein-protein clashes. The electrophoretic mobility shift assay (EMSA) shows that the ETS protein GABPα and the B-ZIP protein CREB preferentially bind to the ETS⇔CRE motif only when the two TFBS overlap precisely. In contrast, the ETS domain of ETV5 and CREB interfere with each other for binding the ETS⇔CRE. The 11-mer (CGGAAGTGACG), the conserved part of the ETS⇔CRE motif, occurs 226 times in the human genome and 83% are in known regulatory regions. In vivo GABPα and CREB ChIP-seq peaks identified the ETS⇔CRE as the most enriched motif occurring in promoters of genes involved in mRNA processing, cellular catabolic processes, and stress response, suggesting that a specific class of genes is regulated by this composite motif. PMID:23050235

  20. Raising the Bar in Reading

    ERIC Educational Resources Information Center

    Naidoo, Taryn; Naicker, Roshilla

    2006-01-01

    This article aims to present strategies and discussion about how teachers can adapt the curriculum in order to provide authentic needs-based programmes. Identifying students' strengths and needs gave us clear directions as to what we needed to teach. In this article, the research team discuss how they successfully raised the reading achievement of…

  1. Raising Boys' Achievement in Schools.

    ERIC Educational Resources Information Center

    Bleach, Kevan, Ed.

    This book offers insights into the range of strategies and good practice being used to raise the achievement of boys. Case studies by school-based practitioners suggest ideas and measures to address the issue of achievement by boys. The contributions are: (1) "Why the Likely Lads Lag Behind" (Kevan Bleach); (2) "Helping Boys Do Better in Their…

  2. Central American Grandparents Raising Grandchildren

    ERIC Educational Resources Information Center

    Fuller-Thomson, Esme; Minkler, Meredith

    2007-01-01

    This study documents the prevalence and profile of Central American grandparents raising grandchildren in the United States of America. Using nationally representative data from the Census 2000 Supplementary Survey, Central American grandparent caregivers are compared with their noncare-giving peers. Results indicate that nationally an estimated…

  3. How (Not) to Raise Money

    ERIC Educational Resources Information Center

    Goeree, Jacob K.; Maasland, Emiel; Onderstal, Sander; Turner, John L.

    2005-01-01

    We show that standard winner-pay auctions are inept fund-raising mechanisms because of the positive externality bidders forgo if they top another's high bid. Revenues are suppressed as a result and remain finite even when bidders value a dollar donated the same as a dollar kept. This problem does not occur in lotteries and all-pay auctions, where…

  4. Optochemogenetics (OCG) allows more precise control of genetic engineering in mice with CreER regulators.

    PubMed

    Lu, Xin; Agasti, Sarit S; Vinegoni, Claudio; Waterman, Peter; DePinho, Ronald A; Weissleder, Ralph

    2012-09-19

    New approaches that allow precise spatiotemporal control of gene expression in model organisms at the single cell level are necessary to better dissect the role of specific genes and cell populations in development, disease, and therapy. Here, we describe a new optochemogenetic switch (OCG switch) to control CreER/loxP-mediated recombination via photoactivatable ("caged") tamoxifen analogues in individual cells in cell culture, organoid culture, and in vivo in adult mice. This approach opens opportunities to more fully exploit existing CreER transgenic mouse strains to achieve more precise temporal- and location-specific regulation of genetic events and gene expression. PMID:22917215

  5. Optochemogenetics (OCG) allows more precise control of genetic engineering in mice with CreER regulators

    PubMed Central

    Lu, Xin; Agasti, Sarit S.; Vinegoni, Claudio; Waterman, Peter; DePinho, Ronald A.; Weissleder, Ralph

    2013-01-01

    New approaches that allow precise spatiotemporal control of gene expression in model organisms at the single cell level are necessary to better dissect the role of specific genes and cell populations in development, disease and therapy. Here, we describe a new optochemogenetic switch (OCG switch) to control CreER/loxP-mediated recombination via photoactivatable (“caged”) tamoxifen analogues in individual cells in cell culture, organoid culture and in vivo in adult mice. This approach opens opportunities to more fully exploit existing CreER transgenic mouse strains to achieve more precise temporal- and location-specific regulation of genetic events and gene expression. PMID:22917215

  6. The Cre/loxP system in Giardia lamblia: genetic manipulations in a binucleate tetraploid protozoan.

    PubMed

    Wampfler, Petra B; Faso, Carmen; Hehl, Adrian B

    2014-07-01

    The bacteriophage-derived Cre/loxP system is a valuable tool that has revolutionised genetic and cell biological research in many organisms. We implemented this system in the intestinal parasite Giardia lamblia, an evolutionarily diverged protozoan whose binucleate and tetraploid genome organisation severely limits the application of reverse genetic approaches. We show that Cre-recombinase is functionally expressed in G. lamblia and demonstrate "recycling" of selectable markers. Providing the means for more complex and versatile genetic modifications, this technique massively increases the scope of functional investigations in G. lamblia and other protozoa with similar limitations with respect to genetic manipulation. PMID:24747534

  7. Targeted transgene insertion into the CHO cell genome using Cre recombinase-incorporating integrase-defective retroviral vectors.

    PubMed

    Kawabe, Yoshinori; Shimomura, Takuya; Huang, Shuohao; Imanishi, Suguru; Ito, Akira; Kamihira, Masamichi

    2016-07-01

    Retroviral vectors have served as efficient gene delivery tools in various biotechnology fields. However, viral DNA is randomly inserted into the genome, which can cause problems, such as insertional mutagenesis and gene silencing. Previously, we reported a site-specific gene integration system, in which a transgene is integrated into a predetermined chromosomal locus of Chinese hamster ovary (CHO) cells using integrase-defective retroviral vectors (IDRVs) and Cre recombinase. In this system, a Cre expression plasmid is transfected into founder cells before retroviral transduction. In practical applications of site-specific gene modification such as for hard-to-transfect cells or for in vivo gene delivery, both the transgene and the Cre protein into retroviral virions should be encapsulate. Here, we generated novel hybrid IDRVs in which viral genome and enzymatically active Cre can be delivered (Cre-IDRVs). Cre-IDRVs encoding marker genes, neomycin resistance and enhanced green fluorescent protein (EGFP), flanked by wild-type and mutated loxP sites were produced using an expression plasmid for a chimeric protein of Cre and retroviral gag-pol. After analyzing the incorporation of the Cre protein into retroviral virions by Western blotting, the Cre-IDRV was infected into founder CHO cells, in which marker genes (hygromycin resistance and red fluorescent protein) flanked with corresponding loxP sites are introduced into the genome. G418-resistant colonies expressing GFP appeared and the site-specific integration of the transgene into the expected chromosomal site was confirmed by PCR and sequencing of amplicons. Moreover, when Cre-IDRV carried a gene expression unit for a recombinant antibody, the recombinant cells in which the antibody expression cassette was integrated in a site-specific manner were generated and the cells produced the recombinant antibody. This method may provide a promising tool to perform site-specific gene modification according to Cre

  8. Strategic Analysis Overview

    NASA Technical Reports Server (NTRS)

    Cirillo, William M.; Earle, Kevin D.; Goodliff, Kandyce E.; Reeves, J. D.; Stromgren, Chel; Andraschko, Mark R.; Merrill, R. Gabe

    2008-01-01

    NASA s Constellation Program employs a strategic analysis methodology in providing an integrated analysis capability of Lunar exploration scenarios and to support strategic decision-making regarding those scenarios. The strategic analysis methodology integrates the assessment of the major contributors to strategic objective satisfaction performance, affordability, and risk and captures the linkages and feedbacks between all three components. Strategic analysis supports strategic decision making by senior management through comparable analysis of alternative strategies, provision of a consistent set of high level value metrics, and the enabling of cost-benefit analysis. The tools developed to implement the strategic analysis methodology are not element design and sizing tools. Rather, these models evaluate strategic performance using predefined elements, imported into a library from expert-driven design/sizing tools or expert analysis. Specific components of the strategic analysis tool set include scenario definition, requirements generation, mission manifesting, scenario lifecycle costing, crew time analysis, objective satisfaction benefit, risk analysis, and probabilistic evaluation. Results from all components of strategic analysis are evaluated a set of pre-defined figures of merit (FOMs). These FOMs capture the high-level strategic characteristics of all scenarios and facilitate direct comparison of options. The strategic analysis methodology that is described in this paper has previously been applied to the Space Shuttle and International Space Station Programs and is now being used to support the development of the baseline Constellation Program lunar architecture. This paper will present an overview of the strategic analysis methodology and will present sample results from the application of the strategic analysis methodology to the Constellation Program lunar architecture.

  9. Status epilepticus stimulates NDEL1 expression via the CREB/CRE pathway in the adult mouse brain.

    PubMed

    Choi, Yun-Sik; Lee, Boyoung; Hansen, Katelin F; Aten, Sydney; Horning, Paul; Wheaton, Kelin L; Impey, Soren; Hoyt, Kari R; Obrietan, Karl

    2016-09-01

    Nuclear distribution element-like 1 (NDEL1/NUDEL) is a mammalian homolog of the Aspergillus nidulans nuclear distribution molecule NudE. NDEL1 plays a critical role in neuronal migration, neurite outgrowth and neuronal positioning during brain development; however within the adult central nervous system, limited information is available regarding NDEL1 expression and functions. Here, the goal was to examine inducible NDEL1 expression in the adult mouse forebrain. Immunolabeling revealed NDEL1 within the forebrain, including the cortex and hippocampus, as well as the midbrain and hypothalamus. Expression was principally localized to perikarya. Using a combination of immunolabeling and RNA seq profiling, we detected a marked and long-lasting upregulation of NDEL1 expression within the hippocampus following a pilocarpine-evoked repetitive seizure paradigm. Chromatin immunoprecipitation (ChIP) analysis identified a cAMP response element-binding protein (CREB) binding site within the CpG island proximal to the NDEL1 gene, and in vivo transgenic repression of CREB led to a marked downregulation of seizure-evoked NDEL1 expression. Together these data indicate that NDEL1 is inducibly expressed in the adult nervous system, and that signaling via the CREB/CRE transcriptional pathway is likely involved. The role of NDEL1 in neuronal migration and neurite outgrowth during development raises the interesting prospect that inducible NDEL1 in the mature nervous system could contribute to the well-characterized structural and functional plasticity resulting from repetitive seizure activity. PMID:27298008

  10. Low cutter load raise head

    SciTech Connect

    Saxman, W.C.

    1981-03-31

    A raise head having a multiplicity of cutters for enlarging a pilot hole into a larger diameter hole by disintegrating the earth formations that surround the pilot hole is provided that will require lower cutter loads to penetrate the formations being bored by directing the rock fracture planes toward the pilot hole forcing the rock to yield with less input energy. The cutters are positioned on the raise head to provide an earth formation contact profile with a major portion of said earth formation contact profile extending outward and upward from said pilot hole. The included angle between the major portion of the earth formation contact profile and the axis of the pilot hole is less than 90/sup 0/.

  11. Physiological characterisation of recombinant Aspergillus nidulans strains with different creA genotypes expressing A. oryzae alpha-amylase.

    PubMed

    Agger, Teit; Petersen, Jesper B; O'Connor, Susan M; Murphy, Rachael L; Kelly, Joan M; Nielsen, Jens

    2002-01-18

    The physiology of three strains of Aspergillus nidulans was examined--a creA deletion strain, a wild type creA genotype and a strain containing extra copies of the creA gene, all producing Aspergillus oryzae alpha-amylase. The strains were cultured in batch and continuous cultivations and the biomass formation and alpha-amylase production was characterised. Overexpression of the creA gene resulted in a lower maximum specific growth rate and a slightly higher repression of the alpha-amylase production during conditions with high glucose concentration. No expression of creA also resulted in a decreased maximum specific growth rate, but also in drastic changes in morphology. Furthermore, the expression of alpha-amylase was completely derepressed and creA thus seems to be the only regulatory protein responsible for glucose repression of alpha-amylase expression. The effect of different carbon sources on the alpha-amylase production in the creA deletion strain was investigated and it was found that starch was the best inducer. The degree of induction by starch increased almost linearly with the concentration of starch in starch/glucose mixtures. High-density batch cultivation was performed with the creA deletion strain and a final titre of 6.0 g l(-1) of alpha-amylase was reached after 162 h of cultivation. PMID:11689252

  12. Official Flag Raising at MTO

    NASA Technical Reports Server (NTRS)

    1962-01-01

    Government officials officially raise the American flag for the first time in front of the Rouchon House in 1962 at the Mississippi Test Operations, denoting NASA's presence in South Mississippi. Pictured, from left, are: Bart Slattery, public affairs officer; Dr. Wernher von Braun, director; Capt. William Fortune, first site manager; Dr. George Constan, manager, Michoud Assembly Facility; Dr. Oswald Lange, chief, Saturn Program Office; Dr. Hermann Weidener, chief, Structures and Mechanics Laboratory; Dr. Karl Heimburg, director, Test Laboratory; and Dan Driscoll, Test Laboratory.

  13. THE CRE-LOXP RECOMBINATION-BASED REPORTER SYSTEM FOR PLANT TRANSCRIPTIONAL EXPRESSION STUDIES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To facilitate the characterization of plant genes, the Cre-loxP site-specific recombination system was adapted to make reporter vectors for plant expression studies. This system allows promoter fragments to be cloned into a small vector (univector) and subsequently recombined in vitro with binary ve...

  14. Cre-Lox Neurogenetics: 20 Years of Versatile Applications in Brain Research and Counting….

    PubMed

    Tsien, Joe Z

    2016-01-01

    Defining and manipulating specific neurons in the brain has garnered enormous interest in recent years, because such an approach is now widely recognized as crucial for deepening our understanding of how the brain works. When I started exploring the Cre-loxP recombination for brain research in the early 1990s, it was written off as a dead-end project by a young fool. Yet over the past 20 years, Cre-lox recombination-mediated neurogenetics has emerged as one of the most powerful and versatile technology platforms for cell-specific gene knockouts, transgenic overexpression, Brainbow imaging, neural pathway tracing with retrovirus and CLARITY, chemical genetics, and optogenetics. Its popularity and greater utility in neuroscience research is also largely thanks to the NIH's bold Blueprint for Neuroscience Research Initiative to launch several Cre-driver resource projects, as well as individual laboratories and private research organizations. With newly-discovered, genetically-encoded molecules that are capable of responding to sonar and magnetic stimulation, for sonogenetics or magnetogenetics, respectively, or detecting rapid voltage changes in neurons, Cre-lox neurogenetics will continue to aid brain research for years to come. PMID:26925095

  15. Genome Editing via Mobile Group-II Introns and Cre/lox

    NASA Astrophysics Data System (ADS)

    Enyeart, P. E.; Perutka, J.; Dao, M.; Ellington, A. E.

    2010-04-01

    Mobile group-II introns and the Cre/lox systems are combined to allow large segments of DNA to be removed or transferred within/between bacterial genomes. Planned applications include metabolic optimization and development of novel dissimilatory metal-reducing bacteria.

  16. CRE-MEDIATED DELETION PRODUCT MAY BE RETAINED AS A CIRCULAR DNA IN TRANSGENIC PLANTS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Earlier we reported using the Cre recombinase to resolve complex multi-copy integration patterns into a single copy. This resolution process also removed the marker gene bar. Four complex-locus wheat lines were obtained by transformation with a lox-plasmid, pVS11. Each of the four lines was crossed ...

  17. Induced Pluripotent Stem Cells Generated from P0-Cre;Z/EG Transgenic Mice

    PubMed Central

    Ogawa, Yasuhiro; Eto, Akira; Miyake, Chisato; Tsuchida, Nana; Miyake, Haruka; Takaku, Yasuhiro; Hagiwara, Hiroaki; Oishi, Kazuhiko

    2015-01-01

    Neural crest (NC) cells are a migratory, multipotent cell population that arises at the neural plate border, and migrate from the dorsal neural tube to their target tissues, where they differentiate into various cell types. Abnormal development of NC cells can result in severe congenital birth defects. Because only a limited number of cells can be obtained from an embryo, mechanistic studies are difficult to perform with directly isolated NC cells. Protein zero (P0) is expressed by migrating NC cells during the early embryonic period. In the P0-Cre;Z/EG transgenic mouse, transient activation of the P0 promoter induces Cre-mediated recombination, indelibly tagging NC-derived cells with enhanced green fluorescent protein (EGFP). Induced pluripotent stem cell (iPSC) technology offers new opportunities for both mechanistic studies and development of stem cell-based therapies. Here, we report the generation of iPSCs from the P0-Cre;Z/EG mouse. P0-Cre;Z/EG mouse-derived iPSCs (P/G-iPSCs) exhibited pluripotent stem cell properties. In lineage-directed differentiation studies, P/G-iPSCs were efficiently differentiated along the neural lineage while expressing EGFP. These results suggest that P/G-iPSCs are useful to study NC development and NC-associated diseases. PMID:26382630

  18. Induced Pluripotent Stem Cells Generated from P0-Cre;Z/EG Transgenic Mice.

    PubMed

    Ogawa, Yasuhiro; Eto, Akira; Miyake, Chisato; Tsuchida, Nana; Miyake, Haruka; Takaku, Yasuhiro; Hagiwara, Hiroaki; Oishi, Kazuhiko

    2015-01-01

    Neural crest (NC) cells are a migratory, multipotent cell population that arises at the neural plate border, and migrate from the dorsal neural tube to their target tissues, where they differentiate into various cell types. Abnormal development of NC cells can result in severe congenital birth defects. Because only a limited number of cells can be obtained from an embryo, mechanistic studies are difficult to perform with directly isolated NC cells. Protein zero (P0) is expressed by migrating NC cells during the early embryonic period. In the P0-Cre;Z/EG transgenic mouse, transient activation of the P0 promoter induces Cre-mediated recombination, indelibly tagging NC-derived cells with enhanced green fluorescent protein (EGFP). Induced pluripotent stem cell (iPSC) technology offers new opportunities for both mechanistic studies and development of stem cell-based therapies. Here, we report the generation of iPSCs from the P0-Cre;Z/EG mouse. P0-Cre;Z/EG mouse-derived iPSCs (P/G-iPSCs) exhibited pluripotent stem cell properties. In lineage-directed differentiation studies, P/G-iPSCs were efficiently differentiated along the neural lineage while expressing EGFP. These results suggest that P/G-iPSCs are useful to study NC development and NC-associated diseases. PMID:26382630

  19. Cre-dependent DNA recombination activates a STING-dependent innate immune response

    PubMed Central

    Pépin, Geneviève; Ferrand, Jonathan; Höning, Klara; Jayasekara, W. Samantha N.; Cain, Jason E.; Behlke, Mark A.; Gough, Daniel J.; G. Williams, Bryan R.; Hornung, Veit; Gantier, Michael P.

    2016-01-01

    Gene-recombinase technologies, such as Cre/loxP-mediated DNA recombination, are important tools in the study of gene function, but have potential side effects due to damaging activity on DNA. Here we show that DNA recombination by Cre instigates a robust antiviral response in mammalian cells, independent of legitimate loxP recombination. This is due to the recruitment of the cytosolic DNA sensor STING, concurrent with Cre-dependent DNA damage and the accumulation of cytoplasmic DNA. Importantly, we establish a direct interplay between this antiviral response and cell–cell interactions, indicating that low cell densities in vitro could be useful to help mitigate these effects of Cre. Taking into account the wide range of interferon stimulated genes that may be induced by the STING pathway, these results have broad implications in fields such as immunology, cancer biology, metabolism and stem cell research. Further, this study sets a precedent in the field of gene-engineering, possibly applicable to other enzymatic-based genome editing technologies. PMID:27166376

  20. Cre-Lox Neurogenetics: 20 Years of Versatile Applications in Brain Research and Counting…

    PubMed Central

    Tsien, Joe Z.

    2016-01-01

    Defining and manipulating specific neurons in the brain has garnered enormous interest in recent years, because such an approach is now widely recognized as crucial for deepening our understanding of how the brain works. When I started exploring the Cre-loxP recombination for brain research in the early 1990s, it was written off as a dead-end project by a young fool. Yet over the past 20 years, Cre-lox recombination-mediated neurogenetics has emerged as one of the most powerful and versatile technology platforms for cell-specific gene knockouts, transgenic overexpression, Brainbow imaging, neural pathway tracing with retrovirus and CLARITY, chemical genetics, and optogenetics. Its popularity and greater utility in neuroscience research is also largely thanks to the NIH’s bold Blueprint for Neuroscience Research Initiative to launch several Cre-driver resource projects, as well as individual laboratories and private research organizations. With newly-discovered, genetically-encoded molecules that are capable of responding to sonar and magnetic stimulation, for sonogenetics or magnetogenetics, respectively, or detecting rapid voltage changes in neurons, Cre-lox neurogenetics will continue to aid brain research for years to come. PMID:26925095

  1. Mutations in the CRE pocket of bacterial RNA polymerase affect multiple steps of transcription

    PubMed Central

    Petushkov, Ivan; Pupov, Danil; Bass, Irina; Kulbachinskiy, Andrey

    2015-01-01

    During transcription, the catalytic core of RNA polymerase (RNAP) must interact with the DNA template with low-sequence specificity to ensure efficient enzyme translocation and RNA extension. Unexpectedly, recent structural studies of bacterial promoter complexes revealed specific interactions between the nontemplate DNA strand at the downstream edge of the transcription bubble (CRE, core recognition element) and a protein pocket formed by core RNAP (CRE pocket). We investigated the roles of these interactions in transcription by analyzing point amino acid substitutions and deletions in Escherichia coli RNAP. The mutations affected multiple steps of transcription, including promoter recognition, RNA elongation and termination. In particular, we showed that interactions of the CRE pocket with a nontemplate guanine immediately downstream of the active center stimulate RNA-hairpin-dependent transcription pausing but not other types of pausing. Thus, conformational changes of the elongation complex induced by nascent RNA can modulate CRE effects on transcription. The results highlight the roles of specific core RNAP–DNA interactions at different steps of RNA synthesis and suggest their importance for transcription regulation in various organisms. PMID:25990734

  2. Cre-dependent DNA recombination activates a STING-dependent innate immune response.

    PubMed

    Pépin, Geneviève; Ferrand, Jonathan; Höning, Klara; Jayasekara, W Samantha N; Cain, Jason E; Behlke, Mark A; Gough, Daniel J; G Williams, Bryan R; Hornung, Veit; Gantier, Michael P

    2016-06-20

    Gene-recombinase technologies, such as Cre/loxP-mediated DNA recombination, are important tools in the study of gene function, but have potential side effects due to damaging activity on DNA. Here we show that DNA recombination by Cre instigates a robust antiviral response in mammalian cells, independent of legitimate loxP recombination. This is due to the recruitment of the cytosolic DNA sensor STING, concurrent with Cre-dependent DNA damage and the accumulation of cytoplasmic DNA. Importantly, we establish a direct interplay between this antiviral response and cell-cell interactions, indicating that low cell densities in vitro could be useful to help mitigate these effects of Cre. Taking into account the wide range of interferon stimulated genes that may be induced by the STING pathway, these results have broad implications in fields such as immunology, cancer biology, metabolism and stem cell research. Further, this study sets a precedent in the field of gene-engineering, possibly applicable to other enzymatic-based genome editing technologies. PMID:27166376

  3. The Cre-Binding Protein Dcreb-a Is Required for Drosophila Embryonic Development

    PubMed Central

    Rose, R. E.; Gallaher, N. M.; Andrew, D. J.; Goodman, R. H.; Smolik, S. M.

    1997-01-01

    We have previously described the cloning of a cyclic AMP response-element (CRE)-binding protein, dCREB-A, in Drosophila melanogaster that is similar to the mammalian CRE-binding protein CREB. dCREB-A is a member of the bZIP family of transcription factors, shows specific binding to the (CRE), and can activate transcription in cell culture. In this report, we describe the gene structure for dCREB-A, protein expression patterns throughout development and the necessary role for this gene in embryogenesis. The 4.5-kb transcript is encoded in six exons that are distributed over 21 kb of DNA. There are seven start sites and no TATA consensus sequences upstream. The dCREB-A protein is expressed in the nuclei of the embryonic salivary gland, proventriculus and stomadeum. Late in embryogenesis, tracheal cell nuclei and specific nuclei within the segments show staining with anti-dCREB-A antibodies. In adult female ovaries, dCREB-A is expressed in the stage 9 through stage 11 follicle cell nuclei. Null mutations of the dCREB-A gene give rise to animals that no longer express dCREB-A protein and die late in embryogenesis before or at hatching. The absolute requirement of dCREB-A for embryogenesis demonstrates a nonredundant function for a CRE-binding protein that will be useful in studying the role of specific signal transduction cascades in development. PMID:9178009

  4. Overview of BioCreAtIvE: critical assessment of information extraction for biology

    PubMed Central

    Hirschman, Lynette; Yeh, Alexander; Blaschke, Christian; Valencia, Alfonso

    2005-01-01

    Background The goal of the first BioCreAtIvE challenge (Critical Assessment of Information Extraction in Biology) was to provide a set of common evaluation tasks to assess the state of the art for text mining applied to biological problems. The results were presented in a workshop held in Granada, Spain March 28–31, 2004. The articles collected in this BMC Bioinformatics supplement entitled "A critical assessment of text mining methods in molecular biology" describe the BioCreAtIvE tasks, systems, results and their independent evaluation. Results BioCreAtIvE focused on two tasks. The first dealt with extraction of gene or protein names from text, and their mapping into standardized gene identifiers for three model organism databases (fly, mouse, yeast). The second task addressed issues of functional annotation, requiring systems to identify specific text passages that supported Gene Ontology annotations for specific proteins, given full text articles. Conclusion The first BioCreAtIvE assessment achieved a high level of international participation (27 groups from 10 countries). The assessment provided state-of-the-art performance results for a basic task (gene name finding and normalization), where the best systems achieved a balanced 80% precision / recall or better, which potentially makes them suitable for real applications in biology. The results for the advanced task (functional annotation from free text) were significantly lower, demonstrating the current limitations of text-mining approaches where knowledge extrapolation and interpretation are required. In addition, an important contribution of BioCreAtIvE has been the creation and release of training and test data sets for both tasks. There are 22 articles in this special issue, including six that provide analyses of results or data quality for the data sets, including a novel inter-annotator consistency assessment for the test set used in task 2. PMID:15960821

  5. Sandia Strategic Plan 1997

    SciTech Connect

    1997-12-01

    Sandia embarked on its first exercise in corporate strategic planning during the winter of 1989. The results of that effort were disseminated with the publication of Strategic Plan 1990. Four years later Sandia conducted their second major planning effort and published Strategic Plan 1994. Sandia`s 1994 planning effort linked very clearly to the Department of Energy`s first strategic plan, Fueling a Competitive Economy. It benefited as well from the leadership of Lockheed Martin Corporation, the management and operating contractor. Lockheed Martin`s corporate success is founded on visionary strategic planning and annual operational planning driven by customer requirements and technology opportunities. In 1996 Sandia conducted another major planning effort that resulted in the development of eight long-term Strategic Objectives. Strategic Plan 1997 differs from its predecessors in that the robust elements of previous efforts have been integrated into one comprehensive body. The changes implemented so far have helped establish a living strategic plan with a stronger business focus and with clear deployment throughout Sandia. The concept of a personal line of sight for all employees to this strategic plan and its objectives, goals, and annual milestones is becoming a reality.

  6. 11. Strategic planning.

    PubMed

    2014-05-01

    There are several types of planning processes and plans, including strategic, operational, tactical, and contingency. For this document, operational planning includes tactical planning. This chapter examines the strategic planning process and includes an introduction into disaster response plans. "A strategic plan is an outline of steps designed with the goals of the entire organisation as a whole in mind, rather than with the goals of specific divisions or departments". Strategic planning includes all measures taken to provide a broad picture of what must be achieved and in which order, including how to organise a system capable of achieving the overall goals. Strategic planning often is done pre-event, based on previous experience and expertise. The strategic planning for disasters converts needs into a strategic plan of action. Strategic plans detail the goals that must be achieved. The process of converting needs into plans has been deconstructed into its components and includes consideration of: (1) disaster response plans; (2) interventions underway or planned; (3) available resources; (4) current status vs. pre-event status; (5) history and experience of the planners; and (6) access to the affected population. These factors are tempered by the local: (a) geography; (b) climate; (c) culture; (d) safety; and (e) practicality. The planning process consumes resources (costs). All plans must be adapted to the actual conditions--things never happen exactly as planned. PMID:24785808

  7. Strategic Risk Assessment

    NASA Technical Reports Server (NTRS)

    Derleth, Jason; Lobia, Marcus

    2009-01-01

    This slide presentation provides an overview of the attempt to develop and demonstrate a methodology for the comparative assessment of risks across the entire portfolio of NASA projects and assets. It includes information about strategic risk identification, normalizing strategic risks, calculation of relative risk score, and implementation options.

  8. Strategic Leadership in Schools

    ERIC Educational Resources Information Center

    Williams, Henry S.; Johnson, Teryl L.

    2013-01-01

    Strategic leadership is built upon traits and actions that encompass the successful execution of all leadership styles. In a world that is rapidly changing, strategic leadership in schools guides school leader through assuring constant improvement process by anticipating future trends and planning for them and noting that plans must be flexible to…

  9. Strategic Investments Overview

    NASA Technical Reports Server (NTRS)

    Comstock, Doug

    2004-01-01

    This viewgraph presentation provides an overview of the organizational hierarchy for strategic management and strategic investments at NASA. The presentation also relates these topics to the budgets it submits to Congress, strategies for space exploration research and development, and systems analysis.

  10. Manage "Human Capital" Strategically

    ERIC Educational Resources Information Center

    Odden, Allan

    2011-01-01

    To strategically manage human capital in education means restructuring the entire human resource system so that schools not only recruit and retain smart and capable individuals, but also manage them in ways that support the strategic directions of the organization. These management practices must be aligned with a district's education improvement…

  11. Developing Strategic Leaders.

    ERIC Educational Resources Information Center

    Carter, Patricia; Terwilliger, Leatha; Alfred, Richard L.; Hartleb, David; Simone, Beverly

    2002-01-01

    Highlights the importance of developing community college leaders capable of demonstrating strategic leadership and responding to the global forces that influence community college education. Discusses the Consortium for Community College Development's Strategic Leadership Forum and its principles, format, content, and early results. (RC)

  12. The Strategic Reader.

    ERIC Educational Resources Information Center

    Devine, James T., Ed.; And Others

    1986-01-01

    To add a foundation to the growing excitement among educators about the central role they play in helping learners become strategic readers, the articles in this thematic journal provide insight into current reading theory and practice. Richard Telfer's article reviews research on strategic reading and clarifies what is meant by the phrase…

  13. A novel Ncr1-Cre mouse reveals the essential role of STAT5 for NK-cell survival and development.

    PubMed

    Eckelhart, Eva; Warsch, Wolfgang; Zebedin, Eva; Simma, Olivia; Stoiber, Dagmar; Kolbe, Thomas; Rülicke, Thomas; Mueller, Mathias; Casanova, Emilio; Sexl, Veronika

    2011-02-01

    We generated a transgenic mouse line that expresses the Cre recombinase under the control of the Ncr1 (p46) promoter. Cre-mediated recombination was tightly restricted to natural killer (NK) cells, as revealed by crossing Ncr1-iCreTg mice to the eGFP-LSLTg reporter strain. Ncr1-iCreTg mice were further used to study NK cell-specific functions of Stat5 (signal transducers and activators of transcription 5) by generating Stat5(f/f) Ncr1-iCreTg animals. Stat5(f/f) Ncr1-iCreTg mice were largely devoid of NK cells in peripheral lymphoid organs. In the bone marrow, NK-cell maturation was abrogated at the NK cell-precursor stage. Moreover, we found that in vitro deletion of Stat5 in interleukin 2-expanded NK cells was incompatible with NK-cell viability. In vivo assays confirmed the complete abrogation of NK cell-mediated tumor control against B16F10-melanoma cells. In contrast, T cell-mediated tumor surveillance against MC38-adenocarcinoma cells was undisturbed. In summary, the results of our study show that STAT5 has a cell-intrinsic role in NK-cell development and that Ncr1-iCreTg mice are a powerful novel tool with which to study NK-cell development, biology, and function. PMID:21127177

  14. Trichoderma reesei CRE1-mediated Carbon Catabolite Repression in Re-sponse to Sophorose Through RNA Sequencing Analysis.

    PubMed

    Antoniêto, Amanda Cristina Campos; de Paula, Renato Graciano; Castro, Lílian Dos Santos; Silva-Rocha, Rafael; Persinoti, Gabriela Felix; Silva, Roberto Nascimento

    2016-04-01

    Carbon catabolite repression (CCR) mediated by CRE1 in Trichoderma reesei emerged as a mechanism by which the fungus could adapt to new environments. In the presence of readily available carbon sources such as glucose, the fungus activates this mechanism and inhibits the production of cellulolytic complex enzymes to avoid unnecessary energy expenditure. CCR has been well described for the growth of T. reesei in cellulose and glucose, however, little is known about this process when the carbon source is sophorose, one of the most potent inducers of cellulase production. Thus, we performed high-throughput RNA sequencing to better understand CCR during cellulase formation in the presence of sophorose, by comparing the mutant ∆cre1 with its parental strain, QM9414. Of the 9129 genes present in the genome of T. reesei, 184 were upregulated and 344 downregulated in the mutant strain ∆cre1 compared to QM9414. Genes belonging to the CAZy database, and those encoding transcription factors and transporters are among the gene classes that were repressed by CRE1 in the presence of sophorose; most were possible indirectly regulated by CRE1. We also observed that CRE1 activity is carbon-dependent. A recent study from our group showed that in cellulose, CRE1 repress different groups of genes when compared to sophorose. CCR differences between these carbon sources may be due to the release of cellodextrins in the cellulose polymer, resulting in different targets of CRE1 in both carbon sources. These results contribute to a better understanding of CRE1-mediated CCR in T. reesei when glucose comes from a potent inducer of cellulase production such as sophorose, which could prove useful in improving cellulase production by the biotechnology sector. PMID:27226768

  15. The Dmp1-SOST Transgene Interacts With and Downregulates the Dmp1-Cre Transgene and the Rosa(Notch) Allele.

    PubMed

    Zanotti, Stefano; Canalis, Ernesto

    2016-05-01

    Activation of Notch1 in osteocytes of Rosa(Notch) mice, where a loxP-flanked STOP cassette and the Nicd coding sequence were targeted to the reverse orientation splice acceptor (Rosa)26 locus, causes osteopetrosis associated with suppressed Sost expression and enhanced Wnt signaling. To determine whether Sost downregulation mediates the effects of Notch activation in osteocytes, Rosa(Notch) mice were crossed with transgenics expressing Cre recombinase or SOST under the control of the dentin matrix protein (Dmp)1 promoter. Dmp1-SOST transgenics displayed vertebral osteopenia and a modest femoral cancellous and cortical bone phenotype, whereas hemizygous Dmp1-Cre transgenics heterozygous for the Rosa(Notch) allele (Dmp1-Cre;Rosa(Notch)) exhibited osteopetrosis. The phenotype of Notch activation in osteocytes was prevented in Dmp1-Cre;Rosa(Notch) mice hemizygous for the Dmp1-SOST transgene. The effect was associated with downregulated Notch signaling and suppressed Dmp1 and Rosa26 expression. To test whether SOST regulates Notch expression in osteocytes, cortical bone cultures from Dmp1-Cre;Rosa(Notch) mice or from Rosa(Notch) control littermates were exposed to recombinant human SOST. The addition of SOST had only modest effects on Notch target gene mRNA levels and suppressed Dmp1, but not Cre or Rosa26, expression. These findings suggest that prevention of the Dmp1-Cre;Rosa(Notch) skeletal phenotype by Dmp1-SOST is not secondary to SOST expression but to interactions among the Dmp1-SOST and Dmp1-Cre transgenes and the Rosa26 locus. In conclusion, the Dmp1-SOST transgene suppresses the expression of the Dmp1-Cre transgene and of Rosa26. PMID:26456319

  16. Partial integration raises antitrust concerns.

    PubMed

    Brock, T H; Kamoie, B E

    2000-11-01

    Recently, providers have begun to explore a new model of integrated delivery system, the partially integrated IDS. Typically, a partially integrated IDS is a joint venture, owned by a core group of providers that maintains complete financial and operational independence outside the joint venture. The IDS contracts with other providers to furnish services that the part-owners do not furnish. A partially integrated IDS raises antitrust concerns because the participating providers may be seen as competitors banding together to set prices jointly for healthcare services. Therefore, to minimize their antitrust exposure, providers that are considering this model should be careful to structure the IDS in accordance with the relevant Federal antitrust laws (i.e., Section 1 of the Sherman Act), taking into account the Federal antitrust agencies' various guidelines and enforcement policies. PMID:11688054

  17. Cre-dependent DREADD (Designer Receptors Exclusively Activated by Designer Drugs) mice.

    PubMed

    Zhu, Hu; Aryal, Dipendra K; Olsen, Reid H J; Urban, Daniel J; Swearingen, Amanda; Forbes, Stacy; Roth, Bryan L; Hochgeschwender, Ute

    2016-08-01

    DREADDs, designer receptors exclusively activated by designer drugs, are engineered G protein-coupled receptors (GPCR) which can precisely control GPCR signaling pathways (for example, Gq, Gs, and Gi). This chemogenetic technology for control of GPCR signaling has been successfully applied in a variety of in vivo studies, including in mice, to remotely control GPCR signaling, for example, in neurons, glia cells, pancreatic β-cells, or cancer cells. In order to fully explore the in vivo applications of the DREADD technology, we generated hM3Dq and hM4Di strains of mice which allow for Cre recombinase-mediated restricted expression of these pathway-selective DREADDs. With the many Cre driver lines now available, these DREADD lines will be applicable to studying a wide array of research and preclinical questions. genesis 54:439-446, 2016. © 2016 Wiley Periodicals, Inc. PMID:27194399

  18. A Specificity Switch in Selected Cre Recombinase Variants Is Mediated by Macromolecular Plasticity and Water

    PubMed Central

    Baldwin, Enoch P.; Martin, Shelley S.; Abel, Jonas; Gelato, Kathy A.; Kim, Hanseong; Schultz, Peter G.; Santoro, Stephen W.

    2010-01-01

    Summary The basis for the altered DNA specificities of two Cre recombinase variants, obtained by mutation and selection, was revealed by their cocrystal structures. The proteins share similar substitutions but differ in their preferences for the natural LoxP substrate and an engineered substrate that is inactive with wild-type Cre, LoxM7. One variant preferentially recombines LoxM7 and contacts the substituted bases through a hydrated network of novel interlocking protein-DNA contacts. The other variant recognizes both LoxP and LoxM7 utilizing the same DNA backbone contact but different base contacts, facilitated by an unexpected DNA shift. Assisted by water, novel interaction networks can arise from few protein substitutions, suggesting how new DNA binding specificities might evolve. The contributions of macromolecular plasticity and water networks in specific DNA recognition observed here present a challenge for predictive schemes. PMID:14652076

  19. A specificity switch in selected cre recombinase variants is mediated by macromolecular plasticity and water.

    PubMed

    Baldwin, Enoch P; Martin, Shelley S; Abel, Jonas; Gelato, Kathy A; Kim, Hanseong; Schultz, Peter G; Santoro, Stephen W

    2003-11-01

    The basis for the altered DNA specificities of two Cre recombinase variants, obtained by mutation and selection, was revealed by their cocrystal structures. The proteins share similar substitutions but differ in their preferences for the natural LoxP substrate and an engineered substrate that is inactive with wild-type Cre, LoxM7. One variant preferentially recombines LoxM7 and contacts the substituted bases through a hydrated network of novel interlocking protein-DNA contacts. The other variant recognizes both LoxP and LoxM7 utilizing the same DNA backbone contact but different base contacts, facilitated by an unexpected DNA shift. Assisted by water, novel interaction networks can arise from few protein substitutions, suggesting how new DNA binding specificities might evolve. The contributions of macromolecular plasticity and water networks in specific DNA recognition observed here present a challenge for predictive schemes. PMID:14652076

  20. Strategic Plan. Volume 1

    NASA Technical Reports Server (NTRS)

    2002-01-01

    The purpose of this document is to present the strategic plan and associated organizational structure that the National Space Biomedical Research Institute (NSBRI) will utilize to achieve the defined mission and objectives provided by NASA. Much of the information regarding the background and establishment of the NSBRI by NASA has been provided in other documentation and will not be repeated in this Strategic Plan. This Strategic Plan is presented in two volumes. Volume I (this volume) begins with an Introduction (Section 2) that provides the Institute's NASA-defined mission and objectives, and the organizational structure adopted to implement these through three Strategic Programs: Countermeasure Research; Education, Training and Outreach; and Cooperative Research and Development. These programs are described in Sections 3 to 5. Each program is presented in a similar way, using four subsections: Goals and Objectives; Current Strategies; Gaps and Modifications; and Resource Requirements. Section 6 provides the administrative infrastructure and total budget required to implement the Strategic Programs and assures that they form a single cohesive plan. This plan will ensure continued success of the Institute for the next five years. Volume II of the Strategic Plan provides an in-depth analysis of the current and future strategic programs of the 12 current NSBRI teams, including their goals, objectives, mutual interactions and schedules.

  1. American Strategic Minerals

    NASA Astrophysics Data System (ADS)

    DeYoung, John H., Jr.; Chidester, Alfred H.

    American Strategic Minerals is a collection of six papers that were presented in December 1982 at a conference organized by the Center for the Study of Marine Policy at the University of Delaware. According to editor Gerard J. Mangone, director of the center, the papers were commissioned “to investigate not only the objective resource situation, but also past United States policy on strategic minerals and future options open to Washington.” The authors and their chapter titles are John C. Kraft, University of Delaware: “Strategic minerals and world stability” V. Anthony Cammarota, Jr., U.S. Bureau of Mines: “America's dependence on strategic minerals” John D. Morgan, U.S. Bureau of Mines: “Future demands of the United States for strategic minerals” J. Robert Moore, University of Texas: “Alternative sources of strategic minerals from the seabed” Allan I. Mendelowitz and John E. Watson, U.S. General Accounting Office: “U.S. mining investments in developing countries” and James W. Curlin, Nautilus Press: “The political dimensions of strategic minerals.”

  2. Epidemiology of carbapenem resistant Enterobacteriaceae (CRE) during 2000-2012 in Asia

    PubMed Central

    Xu, Yanling; Gu, Bing; Huang, Mao; Liu, Haiyan; Xu, Ting; Xia, Wenying

    2015-01-01

    Background Over the past decade, the worldwide emergence of carbapenem resistance in Enterobacteriaceae has become a severe public health issue. This meta-analysis aims to describe the epidemiology of carbapenem resistant Enterobacteriaceae (CRE) during the years of 2000-2012 in Asian area. Methods PubMed and Embase databases were searched to identify the qualified papers. Random or fixed-effect model was used to deal with the data. Results Over all the 49 Asian countries (or regions), only 37.5% [19] of them contributed epidemiology data of CRE, and the rest ones provided either only case reports or no information at all. In Asia, the prevalence of CRE was still low during the study period with average resistance rates of 0.6% (95% CI, 0.6-0.8%, imipenem) and 0.9% (95% CI, 0.7-1.2%, meropenem). Resistance rates to imipenem and meropenem in Enterobacteriaceae exhibited stably escalating trend. Similar trend can also be observed among each Enterobacteriaceae genus, such as E. coli, Klebsiella spp. and Enterobacer spp. Klebsiella spp. accounted for the largest proportion among the isolates resistant to imipenem, and then followed by E. coli and Serratia. The rank order of resistance rates to imipenem among Enterobacteriaceae genus during the period of 2000-2012 was as follows: Serratia spp. (1.8%) > Proteus spp. (1.6%) > Klebsiella spp. (0.8%) = Citrobacter spp. (0.8%) > Enterobacer spp. (0.7%) > E. coli (0.2%). Conclusions Given the fact that the prevalence of CRE was increasing during the past decade, it is urgent for us to establish regional surveillance worldwide, carry out more effective antibiotic stewardship and infection control measures to prevent further spread of carbapenem resistance in Enterobacteriaceae. PMID:25922715

  3. Long-term Cre-mediated retrograde tagging of neurons using a novel recombinant pseudorabies virus

    PubMed Central

    Oyibo, Hassana K.; Znamenskiy, Petr; Oviedo, Hysell V.; Enquist, Lynn W.; Zador, Anthony M.

    2014-01-01

    Brain regions contain diverse populations of neurons that project to different long-range targets. The study of these subpopulations in circuit function and behavior requires a toolkit to characterize and manipulate their activity in vivo. We have developed a novel set of reagents based on Pseudorabies Virus (PRV) for efficient and long-term genetic tagging of neurons based on their projection targets. By deleting IE180, the master transcriptional regulator in the PRV genome, we have produced a mutant virus capable of infection and transgene expression in neurons but unable to replicate in or spread from those neurons. IE180-null mutants showed no cytotoxicity, and infected neurons exhibited normal physiological function more than 45 days after infection, indicating the utility of these engineered viruses for chronic experiments. To enable rapid and convenient construction of novel IE180-null recombinants, we engineered a bacterial artificial chromosome (BAC) shuttle-vector system for moving new constructs into the PRV IE180-null genome. Using this system we generated an IE180-null recombinant virus expressing the site-specific recombinase Cre. This Cre-expressing virus (PRV-hSyn-Cre) efficiently and robustly infects neurons in vivo and activates transgene expression from Cre-dependent vectors in local and retrograde projecting populations of neurons in the mouse. We also generated an assortment of recombinant viruses expressing fluorescent proteins (mCherry, EGFP, ECFP). These viruses exhibit long-term labeling of neurons in vitro but transient labeling in vivo. Together these novel IE180-null PRV reagents expand the toolkit for targeted gene expression in the brain, facilitating functional dissection of neuronal circuits in vivo. PMID:25232307

  4. Characterization of Channelrhodopsin and Archaerhodopsin in Cholinergic Neurons of Cre-Lox Transgenic Mice

    PubMed Central

    Hedrick, Tristan; Danskin, Bethanny; Larsen, Rylan S.; Ollerenshaw, Doug; Groblewski, Peter; Valley, Matthew; Olsen, Shawn; Waters, Jack

    2016-01-01

    The study of cholinergic signaling in the mammalian CNS has been greatly facilitated by the advent of mouse lines that permit the expression of reporter proteins, such as opsins, in cholinergic neurons. However, the expression of opsins could potentially perturb the physiology of opsin-expressing cholinergic neurons or mouse behavior. Indeed, the published literature includes examples of cellular and behavioral perturbations in preparations designed to drive expression of opsins in cholinergic neurons. Here we investigate expression of opsins, cellular physiology of cholinergic neurons and behavior in two mouse lines, in which channelrhodopsin-2 (ChR2) and archaerhodopsin (Arch) are expressed in cholinergic neurons using the Cre-lox system. The two mouse lines were generated by crossing ChAT-Cre mice with Cre-dependent reporter lines Ai32(ChR2-YFP) and Ai35(Arch-GFP). In most mice from these crosses, we observed expression of ChR2 and Arch in only cholinergic neurons in the basal forebrain and in other putative cholinergic neurons in the forebrain. In small numbers of mice, off-target expression occurred, in which fluorescence did not appear limited to cholinergic neurons. Whole-cell recordings from fluorescently-labeled basal forebrain neurons revealed that both proteins were functional, driving depolarization (ChR2) or hyperpolarization (Arch) upon illumination, with little effect on passive membrane properties, spiking pattern or spike waveform. Finally, performance on a behavioral discrimination task was comparable to that of wild-type mice. Our results indicate that ChAT-Cre x reporter line crosses provide a simple, effective resource for driving indicator and opsin expression in cholinergic neurons with few adverse consequences and are therefore an valuable resource for studying the cholinergic system. PMID:27243816

  5. Utilizing the CreA Gene to Better Understand Carbon Flux in Fungi

    SciTech Connect

    Annecharico, Matthew D.; Baker, Scott E.

    2005-01-01

    Carbon catabolite repression (CCR) is a system which represses the synthesis of enzymes required for certain carbon sources when more favored carbon sources are present. Fungal enzymes are useful in various industrial applications such as the degradation of bi-product biomass produced in many areas of manufacturing and production. For example, the basidiomycete, Phanerochaete chrysosporium, a filamentous fungus, produces a number of lignin peroxidases (LiPs) such as an extra-cellular lignin-modifying protein and manganese-dependent peroxidase (MnPs). The LiPs secreted by P. chrysosporium are able to break down lignin, one of the main substances of wood, while leaving the cellulose behind. These enzymes are capable of degrading toxic waste as well as biomass waste. Through polysaccharide hydrolysis and lignin utilization, biobased products and biofuels can be processed from biomass instead of petroleum, which is more environmentally sound than current fuel options. Previous research in the fungal phylum, Ascomyces has shown that the creA gene is an important gene that controls various enzymes which have key roles in controlling carbon utilization by fungi. We hypothesize that there are similarities across all fungi in the mechanism by which carbon flux is controlled and that orthologs of the creA gene exist in all fungal phyla. A COnsensus-DEgenerate Hybrid Oligonucleotide Primers (CODEHOP) polymerase chain reaction (PCR) strategy was used to isolate genomic fragments of the creA gene from a variety of fungi with the objective to design and utilize a method for isolation of the creA gene from a wide range of fungi.

  6. Being a Learner Using Social Media in School: The Case of Space2cre8

    ERIC Educational Resources Information Center

    Vasbø, Kristin Beate; Silseth, Kenneth; Erstad, Ola

    2014-01-01

    The aim of this article is to gain knowledge about what it means to be a learner using social media in an educational setting. The article presents an ethnographic study of students in a multiethnic community in Oslo who participate in a social networking site called Space2cre8 (S28). In this article, we set out to explore the kind of space for…

  7. Successful Processes to Engender Board Ownership of Strategic Planning

    ERIC Educational Resources Information Center

    Calareso, Jack P.

    2007-01-01

    Too often, Boards of Trustees feel that final approval of the institution's Strategic Plan is sufficient. However, most Plans will only be successfully implemented if the Board has complete ownership and a commitment to be actively involved in many areas of the institution (not only fund raising). This article describes a successful process and…

  8. California Community Colleges Technology II Strategic Plan, 2000-2005.

    ERIC Educational Resources Information Center

    California Community Colleges, Sacramento. Office of the Chancellor.

    This Strategic Plan aims to address the issues facing California Community Colleges (CCCs) due to new technology. The issues raised by the report include: (1) the pervasive and economically and socially necessary use of the Internet; (2) the digital divide; (3) the need for integrating technology into teaching and learning; (4) the impact of Tidal…

  9. Generation and Characterisation of a Pax8-CreERT2 Transgenic Line and a Slc22a6-CreERT2 Knock-In Line for Inducible and Specific Genetic Manipulation of Renal Tubular Epithelial Cells

    PubMed Central

    Espana-Agusti, Judit; Zou, Xiangang; Wong, Kim; Fu, Beiyuan; Yang, Fengtang; Tuveson, David A.; Adams, David J.; Matakidou, Athena

    2016-01-01

    Genetically relevant mouse models need to recapitulate the hallmarks of human disease by permitting spatiotemporal gene targeting. This is especially important for replicating the biology of complex diseases like cancer, where genetic events occur in a sporadic fashion within developed somatic tissues. Though a number of renal tubule targeting mouse lines have been developed their utility for the study of renal disease is limited by lack of inducibility and specificity. In this study we describe the generation and characterisation of two novel mouse lines directing CreERT2 expression to renal tubular epithelia. The Pax8-CreERT2 transgenic line uses the mouse Pax8 promoter to direct expression of CreERT2 to all renal tubular compartments (proximal and distal tubules as well as collecting ducts) whilst the Slc22a6-CreERT2 knock-in line utilises the endogenous mouse Slc22a6 locus to specifically target the epithelium of proximal renal tubules. Both lines show high organ and tissue specificity with no extrarenal activity detected. To establish the utility of these lines for the study of renal cancer biology, Pax8-CreERT2 and Slc22a6-CreERT2 mice were crossed to conditional Vhl knockout mice to induce long-term renal tubule specific Vhl deletion. These models exhibited renal specific activation of the hypoxia inducible factor pathway (a VHL target). Our results establish Pax8-CreERT2 and Slc22a6-CreERT2 mice as valuable tools for the investigation and modelling of complex renal biology and disease. PMID:26866916

  10. Generation and Characterisation of a Pax8-CreERT2 Transgenic Line and a Slc22a6-CreERT2 Knock-In Line for Inducible and Specific Genetic Manipulation of Renal Tubular Epithelial Cells.

    PubMed

    Espana-Agusti, Judit; Zou, Xiangang; Wong, Kim; Fu, Beiyuan; Yang, Fengtang; Tuveson, David A; Adams, David J; Matakidou, Athena

    2016-01-01

    Genetically relevant mouse models need to recapitulate the hallmarks of human disease by permitting spatiotemporal gene targeting. This is especially important for replicating the biology of complex diseases like cancer, where genetic events occur in a sporadic fashion within developed somatic tissues. Though a number of renal tubule targeting mouse lines have been developed their utility for the study of renal disease is limited by lack of inducibility and specificity. In this study we describe the generation and characterisation of two novel mouse lines directing CreERT2 expression to renal tubular epithelia. The Pax8-CreERT2 transgenic line uses the mouse Pax8 promoter to direct expression of CreERT2 to all renal tubular compartments (proximal and distal tubules as well as collecting ducts) whilst the Slc22a6-CreERT2 knock-in line utilises the endogenous mouse Slc22a6 locus to specifically target the epithelium of proximal renal tubules. Both lines show high organ and tissue specificity with no extrarenal activity detected. To establish the utility of these lines for the study of renal cancer biology, Pax8-CreERT2 and Slc22a6-CreERT2 mice were crossed to conditional Vhl knockout mice to induce long-term renal tubule specific Vhl deletion. These models exhibited renal specific activation of the hypoxia inducible factor pathway (a VHL target). Our results establish Pax8-CreERT2 and Slc22a6-CreERT2 mice as valuable tools for the investigation and modelling of complex renal biology and disease. PMID:26866916

  11. CollagenVI-Cre mice: A new tool to target stromal cells in secondary lymphoid organs.

    PubMed

    Prados, Alejandro; Kollias, George; Koliaraki, Vasiliki

    2016-01-01

    Stromal cells in secondary lymphoid organs (SLOs) are non-hematopoietic cells involved in the regulation of adaptive immune responses. Three major stromal populations have been identified in adult SLOs: fibroblastic reticular cells (FRCs), follicular dendritic cells (FDCs) and marginal reticular cells (MRCs). The properties of these individual populations are not clearly defined, mainly due to the lack of appropriate genetic tools, especially for MRCs. Here, we analyzed stromal cell targeting in SLOs from a transgenic mouse strain that expresses Cre recombinase under the CollagenVI promoter, using lineage tracing approaches. We show that these mice target specifically MRCs and FDCs, but not FRCs in Peyer's patches and isolated lymphoid follicles in the intestine. In contrast, stromal cells in lymph nodes and the spleen do not express the transgene, which renders ColVI-cre mice ideal for the specific targeting of stromal cells in the gut-associated lymphoid tissue (GALT). This funding further supports the hypothesis of organ-specific stromal precursors in SLOs. Interestingly, in all tissues analyzed, there was also high specificity for perivascular cells, which have been proposed to act as FDC precursors. Taken together, ColVI-Cre mice are a useful new tool for the dissection of MRC- and FDC-specific functions and plasticity in the GALT. PMID:27604178

  12. Fabp4-CreER lineage tracing revealstwo distinctive coronary vascular populations

    PubMed Central

    He, Lingjuan; Tian, Xueying; Zhang, Hui; Wythe, Joshua D; Zhou, Bin

    2014-01-01

    Over the last two decades, genetic lineage tracing has allowed for the elucidation of the cellular origins and fates during both embryogenesis and in pathological settings in adults. Recent lineage tracing studies using Apln-CreER tool indicated that a large number of post-natal coronary vessels do not form from pre-existing vessels. Instead, they form de novo after birth, which represents a coronary vascular population (CVP) distinct from the pre-existing one. Herein, we present new coronary vasculature lineage tracing results using a novel tool, Fabp4-CreER. Our results confirm the distinct existence of two unique CVPs. The 1st CVP, which is labelled by Fabp4-CreER, arises through angiogenic sprouting of pre-existing vessels established during early embryogenesis. The 2nd CVP is not labelled by Fabp4, suggesting that these vessels form de novo, rather than through expansion of the 1st CVP. These results support the de novo formation of vessels in the post-natal heart, which has implications for studies in cardiovascular disease and heart regeneration. PMID:25265869

  13. Engineering mouse chromosomes with Cre-loxP: range, efficiency, and somatic applications.

    PubMed

    Zheng, B; Sage, M; Sheppeard, E A; Jurecic, V; Bradley, A

    2000-01-01

    Chromosomal rearrangements are important resources for genetic studies. Recently, a Cre-loxP-based method to introduce defined chromosomal rearrangements (deletions, duplications, and inversions) into the mouse genome (chromosome engineering) has been established. To explore the limits of this technology systematically, we have evaluated this strategy on mouse chromosome 11. Although the efficiency of Cre-loxP-mediated recombination decreases with increasing genetic distance when the two endpoints are on the same chromosome, the efficiency is not limiting even when the genetic distance is maximized. Rearrangements encompassing up to three quarters of chromosome 11 have been constructed in mouse embryonic stem (ES) cells. While larger deletions may lead to ES cell lethality, smaller deletions can be produced very efficiently both in ES cells and in vivo in a tissue- or cell-type-specific manner. We conclude that any chromosomal rearrangement can be made in ES cells with the Cre-loxP strategy provided that it does not affect cell viability. In vivo chromosome engineering can be potentially used to achieve somatic losses of heterozygosity in creating mouse models of human cancers. PMID:10611243

  14. Atom probe field ion microscopy of type 308 CRE stainless steel welds

    SciTech Connect

    Babu, S.S.; David, S.A.; Vitek, J.M.; Miller, M.K.

    1994-09-01

    Additions of controlled residual elements (CRE) such as titanium, boron and phosphorus, to the commercial 308 type stainless steel welds have been found to improve the creep-rupture properties. In both conventional and CRE-type stainless steel welds, during high temperature service, various microstructural changes take place such as the formation of M{sub 23}C{sub 6} carbide networks along austenite({gamma})-ferrite ({delta}) interface, the formation of sigma ({sigma}) phase, and the dissolution of ferrite. Boron-containing CRE welds exhibit similar microstructural evolution as that of conventional types 308 stainless steel welds. As boron may segregate to ferrite-austenite interface and thereby modify the austenite-M{sub 23}C{sub 6} carbide interface, the distribution of boron was measured with an atom probe field ion microscope. Atom probe analysis revealed significant boron and carbon enrichments along the ferrite-austenite boundary in the as-welded state. The ratio of carbon to boron concentration at the interface was found to be {approximately} 4.

  15. CollagenVI-Cre mice: A new tool to target stromal cells in secondary lymphoid organs

    PubMed Central

    Prados, Alejandro; Kollias, George; Koliaraki, Vasiliki

    2016-01-01

    Stromal cells in secondary lymphoid organs (SLOs) are non-hematopoietic cells involved in the regulation of adaptive immune responses. Three major stromal populations have been identified in adult SLOs: fibroblastic reticular cells (FRCs), follicular dendritic cells (FDCs) and marginal reticular cells (MRCs). The properties of these individual populations are not clearly defined, mainly due to the lack of appropriate genetic tools, especially for MRCs. Here, we analyzed stromal cell targeting in SLOs from a transgenic mouse strain that expresses Cre recombinase under the CollagenVI promoter, using lineage tracing approaches. We show that these mice target specifically MRCs and FDCs, but not FRCs in Peyer’s patches and isolated lymphoid follicles in the intestine. In contrast, stromal cells in lymph nodes and the spleen do not express the transgene, which renders ColVI-cre mice ideal for the specific targeting of stromal cells in the gut-associated lymphoid tissue (GALT). This funding further supports the hypothesis of organ-specific stromal precursors in SLOs. Interestingly, in all tissues analyzed, there was also high specificity for perivascular cells, which have been proposed to act as FDC precursors. Taken together, ColVI-Cre mice are a useful new tool for the dissection of MRC- and FDC-specific functions and plasticity in the GALT. PMID:27604178

  16. In vitro susceptibility of β-lactamase-producing carbapenem-resistant Enterobacteriaceae (CRE) to eravacycline.

    PubMed

    Zhang, Yunliang; Lin, Xiaoyan; Bush, Karen

    2016-08-01

    Eravacycline is a novel, fully synthetic fluorocycline antibiotic of the tetracycline class being developed for the treatment of complicated urinary tract infections and complicated intra-abdominal infections. Eravacycline has activity against many key Gram-negative pathogens, including Enterobacteriaceae resistant to carbapenems, cephalosporins, fluoroquinolones and β-lactam/β-lactamase inhibitor combinations, including strains that are multidrug-resistant. Carbapenem-resistant Enterobacteriaceae (CRE) isolates from 2010 to 2013 (n=110) were characterized for carbapenemase genes by PCR and sequencing. MICs for eravacycline, tetracycline, tigecycline, amikacin, imipenem, ceftazidime, cefotaxime and levofloxacin were determined in broth microdilution assays. All isolates produced at least one carbapenemase, most frequently KPC-3. Nine isolates produced both a KPC serine carbapenemase and a metallo-β-lactamase, NDM-1 (n=1) or VIM-1 (n=8). The 110 isolates were highly resistant to all the β-lactams tested and to levofloxacin, and had MIC50/MIC90 values in the intermediate range for tetracycline and amikacin. MIC50/MIC90 values for eravacycline were 1/2 μg ml(-1) compared with 2/2 μg ml(-1) for tigecycline. Eravacycline MICs were often twofold lower than for tigecycline, with 64% of the eravacycline MICs <2 μg ml(-1) as compared with <4% of tigecycline MICs. Overall, eravacycline demonstrated the lowest cumulative MICs against this panel of recent CRE and may have the potential to treat infections caused by CRE. PMID:27353166

  17. Generation of a cre recombinase-conditional Nos1ap over-expression transgenic mouse

    PubMed Central

    Auer, Dallas R.; Sysa-Shah, Polina; Bedja, Djahida; Simmers, Jessica L.; Pak, Evgenia; Dutra, Amalia; Cohn, Ronald; Gabrielson, Kathleen L.

    2016-01-01

    Polymorphic non-coding variants at the NOS1AP locus have been associated with the common cardiac, metabolic and neurological traits and diseases. Although, in vitro gene targeting-based cellular and biochemical studies have shed some light on NOS1AP function in cardiac and neuronal tissue, to enhance our understanding of NOS1AP function in mammalian physiology and disease, we report the generation of cre recombinase-conditional Nos1ap over-expression transgenic mice (Nos1apTg). Conditional transgenic mice were generated by the pronuclear injection method and three independent, single-site, multiple copies integration event-based founder lines were selected. For heart-restricted over-expression, Nos1apTg mice were crossed with Mlc2v-cre and Nos1ap transcript over-expression was observed in left ventricles from Nos1apTg; Mlc2v-cre F1 mice. We believe that with the potential of conditional over-expression, Nos1apTg mice will be a useful resource in studying NOS1AP function in various tissues under physiological and disease states. PMID:24563304

  18. All-in-one construct for genome engineering using Cre-lox technology

    PubMed Central

    Mariscal, Ana M.; González-González, Luis; Querol, Enrique; Piñol, Jaume

    2016-01-01

    Mycoplasma genitalium is an appealing model of a minimal cell and synthetic biology study, and it was one of the first organisms whose genome was fully sequenced and chemically synthesized. Despite its usefulness as a model organism, many genetic tools well established for other microorganisms are not currently available in mycoplasmas. We have developed several vectors to adapt the Cre-lox technology for genome engineering in M. genitalium, providing an all-in-one construct that could be also useful to obtain unmarked genetic modifications in many other slow growing microorganisms. This construct contains a modified promoter sequence based in TetR system that exhibits an enhanced control on Cre recombinase expression, virtually abolishing the presence of this recombinase in the absence of inducer. This allows to introduce the Cre recombinase gene and the desired genetic modification in a single transformation step. In addition, this inducible promoter may be a very promising tool for a wide range of molecular applications. PMID:27084897

  19. Generating mouse models of degenerative diseases using Cre/lox-mediated in vivo mosaic cell ablation

    PubMed Central

    Fujioka, Masato; Tokano, Hisashi; Fujioka, Keiko Shiina; Okano, Hideyuki; Edge, Albert S.B.

    2011-01-01

    Most degenerative diseases begin with a gradual loss of specific cell types before reaching a threshold for symptomatic onset. However, the endogenous regenerative capacities of different tissues are difficult to study, because of the limitations of models for early stages of cell loss. Therefore, we generated a transgenic mouse line (Mos-iCsp3) in which a lox-mismatched Cre/lox cassette can be activated to produce a drug-regulated dimerizable caspase-3. Tissue-restricted Cre expression yielded stochastic Casp3 expression, randomly ablating a subset of specific cell types in a defined domain. The limited and mosaic cell loss led to distinct responses in 3 different tissues targeted using respective Cre mice: reversible, impaired glucose tolerance with normoglycemia in pancreatic β cells; wound healing and irreversible hair loss in the skin; and permanent moderate deafness due to the loss of auditory hair cells in the inner ear. These mice will be important for assessing the repair capacities of tissues and the potential effectiveness of new regenerative therapies. PMID:21576819

  20. Mice expressing GFP and CreER in osteochondro progenitor cells in the periosteum

    PubMed Central

    Kawanami, Aya; Matsushita, Takehiko; Chan, Yuk Yu; Murakami, Shunichi

    2009-01-01

    We generated Prx1CreER-GFP transgenic mice that express tamoxifen-inducible Cre recombinase and GFP under the control of a 2.4 kb Prx1 promoter. The transgene is expressed in osteochondro progenitor cells in the developing limb buds and in a subpopulation of periosteal cells that is closely associated with the cortical bone. GFP-expressing cells isolated from the diaphyses of long bones by cell sorting express multiple markers of periosteal cells, including Prx-1, Fgf18, Tenascin-W, Periostin, and Thrombospondin 2. In addition, these cells undergo chondrogenic and osteogenic differentiation in culture upon induction. Cell fate analysis using the Rosa26 LacZ reporter indicated that transgene-expressing cells give rise to some of the chondrocytes and osteoblasts in the fracture callus. Collectively, these observations strongly suggest that the transgene-expressing cells are osteochondro progenitor cells in the periosteum. The established Prx1CreER-GFP mice would offer novel approaches for analyzing the functions of periosteal cells in vitro and in vivo. PMID:19538944

  1. Cre recombinase expression or topical tamoxifen treatment do not affect retinal structure and function, neuronal vulnerability or glial reactivity in the mouse eye.

    PubMed

    Boneva, S K; Groß, T R; Schlecht, A; Schmitt, S I; Sippl, C; Jägle, H; Volz, C; Neueder, A; Tamm, E R; Braunger, B M

    2016-06-14

    Mice with a constitutive or tamoxifen-induced Cre recombinase (Cre) expression are frequently used research tools to allow the conditional deletion of target genes via the Cre-loxP system. Here we analyzed for the first time in a comprehensive and comparative way, whether retinal Cre expression or topical tamoxifen treatment itself would cause structural or functional changes, including changes in the expression profiles of molecular markers, glial reactivity and photoreceptor vulnerability. To this end, we characterized the transgenic α-Cre, Lmop-Cre and the tamoxifen-inducible CAGG-CreER™ mouse lines, all having robust Cre expression in the neuronal retina. In addition, we characterized the effects of topical tamoxifen treatment itself in wildtype mice. We performed morphometric analyses, immunohistochemical staining, in vivo ERG and angiography analyses and realtime RT-PCR analyses. Furthermore, the influence of Cre recombinase or topical tamoxifen exposure on neuronal vulnerability was studied by using light damage as a model for photoreceptor degeneration. Taken together, neither the expression of Cre, nor topical tamoxifen treatment caused detectable changes in retinal structure and function, the expression profiles of investigated molecular markers, glial reactivity and photoreceptor vulnerability. We conclude that the Cre-loxP system and its induction through tamoxifen is a safe and reliable method to delete desired target genes in the neural retina. PMID:27026593

  2. 7 CFR 60.123 - Raised.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 3 2013-01-01 2013-01-01 false Raised. 60.123 Section 60.123 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards, Inspections... AND SHELLFISH General Provisions Definitions § 60.123 Raised. Raised means in the case of: (a)...

  3. 7 CFR 60.123 - Raised.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 3 2014-01-01 2014-01-01 false Raised. 60.123 Section 60.123 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards, Inspections... AND SHELLFISH General Provisions Definitions § 60.123 Raised. Raised means in the case of: (a)...

  4. Generation of Oxtr cDNA(HA)-Ires-Cre Mice for Gene Expression in an Oxytocin Receptor Specific Manner.

    PubMed

    Hidema, Shizu; Fukuda, Tomokazu; Hiraoka, Yuichi; Mizukami, Hiroaki; Hayashi, Ryotaro; Otsuka, Ayano; Suzuki, Shingo; Miyazaki, Shinji; Nishimori, Katsuhiko

    2016-05-01

    The neurohypophysial hormone oxytocin (OXT) and its receptor (OXTR) have critical roles in the regulation of pro-social behaviors, including social recognition, pair bonding, parental behavior, and stress-related responses. Supporting this hypothesis, a portion of patients suffering from autism spectrum disorder have mutations, such as single nucleotide polymorphisms, or epigenetic modifications in their OXTR gene. We previously reported that OXTR-deficient mice exhibit pervasive social deficits, indicating the critical role of OXTR in social behaviors. In the present study, we generated Oxtr cDNA(HA)-Ires-Cre knock-in mice, expressing both OXTR and Cre recombinase under the control of the endogenous Oxtr promoter. Knock-in cassette of Oxtr cDNA(HA)-Ires-Cre consisted of Oxtr cDNA tagged with the hemagglutinin epitope at the 3' end (Oxtr cDNA(HA)), internal ribosomal entry site (Ires), and Cre. Cre was expressed in the uterus, mammary gland, kidney, and brain of Oxtr cDNA(HA)-Ires-Cre knock-in mice. Furthermore, the distribution of Cre in the brain was similar to that observed in Oxtr-Venus fluorescent protein expressing mice (Oxtr-Venus), another animal model previously generated by our group. Social behavior of Oxtr cDNA(HA)-Ires-Cre knock-in mice was similar to that of wild-type animals. We demonstrated that this construct is expressed in OXTR-expressing neurons specifically after an infection with the recombinant adeno-associated virus carrying the flip-excision switch vector. Using this system, we showed the transport of the wheat-germ agglutinin tracing molecule from the OXTR-expressing neurons to the innervated neurons in knock-in mice. This study might contribute to the monosynaptic analysis of neuronal circuits and to the optogenetic analysis of neurons expressing OXTR. PMID:26442453

  5. The strategic security officer.

    PubMed

    Hodges, Charles

    2014-01-01

    This article discusses the concept of the strategic security officer, and the potential that it brings to the healthcare security operational environment. The author believes that training and development, along with strict hiring practices, can enable a security department to reach a new level of professionalism, proficiency and efficiency. The strategic officer for healthcare security is adapted from the "strategic corporal" concept of US Marine Corps General Charles C. Krulak which focuses on understanding the total force implications of the decisions made by the lowest level leaders within the Corps (Krulak, 1999). This article focuses on the strategic organizational implications of every security officer's decisions in the constantly changing and increasingly volatile operational environment of healthcare security. PMID:24707753

  6. Strategic EDP Planning.

    ERIC Educational Resources Information Center

    Wold, Geoffrey H.

    1988-01-01

    School districts need a strategic, comprehensive, electronic data processing resources plan that would include the following major elements: (1) systems; (2) hardware; (3) software; (4) staffing; (5) control; and (6) projected budgets. (MLF)

  7. 2015 Enterprise Strategic Vision

    SciTech Connect

    2015-08-01

    This document aligns with the Department of Energy Strategic Plan for 2014-2018 and provides a framework for integrating our missions and direction for pursuing DOE’s strategic goals. The vision is a guide to advancing world-class science and engineering, supporting our people, modernizing our infrastructure, and developing a management culture that operates a safe and secure enterprise in an efficient manner.

  8. Site-specific modification of genome with cell-permeable Cre fusion protein in preimplantation mouse embryo

    SciTech Connect

    Kim, Kyoungmi; Kim, Hwain; Lee, Daekee

    2009-10-09

    Site-specific recombination (SSR) by Cre recombinase and its target sequence, loxP, is a valuable tool in genetic analysis of gene function. Recently, several studies reported successful application of Cre fusion protein containing protein transduction peptide for inducing gene modification in various mammalian cells including ES cell as well as in the whole animal. In this study, we show that a short incubation of preimplantation mouse embryos with purified cell-permeable Cre fusion protein results in efficient SSR. X-Gal staining of preimplantation embryos, heterozygous for Gtrosa26{sup tm1Sor}, revealed that treatment of 1-cell or 2-cell embryos with 3 {mu}M of Cre fusion protein for 2 h leads to Cre-mediated excision in 70-85% of embryos. We have examined the effect of the concentration of the Cre fusion protein and the duration of the treatment on embryonic development, established a condition for full term development and survival to adulthood, and demonstrated the germ line transmission of excised Gtrosa26 allele. Potential applications and advantages of the highly efficient technique described here are discussed.

  9. Comparative analysis of the efficiency and specificity of myeloid-Cre deleting strains using ROSA-EYFP reporter mice.

    PubMed

    Abram, Clare L; Roberge, Gray L; Hu, Yongmei; Lowell, Clifford A

    2014-06-01

    Since the first example of conditional gene targeting in mice in 1994, the use of Cre recombinase and loxP flanked sequences has become an invaluable technique to generate tissue and temporal specific gene knockouts. The number of mouse strains expressing floxed-gene sequences, and tissue-specific or temporal-specific Cre-recombinase that have been reported in the literature has grown exponentially. However, increased use of this technology has highlighted several problems that can impact the interpretation of any phenotype observed in these mouse models. In particular, accurate knowledge of the specificity of Cre expression in each strain is critical in order to make conclusions about the role of specific cell types in the phenotypes observed. Cre-mediated deletion specificity and efficiency have been described in many different ways in the literature, making direct comparisons between these Cre strains impossible. Here we report crossing thirteen different myeloid-Cre mouse strains to ROSA-EYFP reporter mice and assaying YFP expression in a variety of naïve unstimulated hematopoietic cells, in parallel. By focusing on myeloid subsets, we directly compare the relative efficiency and specificity of myeloid deletion in these strains under steady-state conditions. PMID:24857755

  10. Comparative analysis of the efficiency and specificity of myeloid-Cre deleting strains using ROSA-EYFP reporter mice

    PubMed Central

    Abram, Clare L.; Roberge, Gray L.; Hu, Yongmei; Lowell, Clifford A.

    2014-01-01

    Since the first example of conditional gene targeting in mice in 1994, the use of Cre recombinase and loxP flanked sequences has become an invaluable technique to generate tissue and temporal specific gene knockouts. The number of mouse strains expressing floxed-gene sequences, and tissue-specific or temporal-specific Cre-recombinase that have been reported in the literature has grown exponentially. However, increased use of this technology has highlighted several problems that can impact the interpretation of any phenotype observed in these mouse models. In particular, accurate knowledge of the specificity of Cre expression in each strain is critical in order to make conclusions about the role of specific cell types in the phenotypes observed. Cre-mediated deletion specificity and efficiency has been described in many different ways in the literature, making direct comparisons between these Cre strains impossible. Here we report crossing thirteen different myeloid-Cre mouse strains to ROSA-EYFP reporter mice and assaying YFP expression in a variety of naïve unstimulated hematopoietic cells, in parallel. By focusing on myeloid subsets, we directly compare the relative efficiency and specificity of myeloid deletion in these strains under steady-state conditions. PMID:24857755

  11. Management of raised intracranial pressure.

    PubMed Central

    Pickard, J D; Czosnyka, M

    1993-01-01

    based on experience of the particular cause of raised intracranial pressure in a given age group. PMID:8350099

  12. Guidelines for strategic planning

    SciTech Connect

    Not Available

    1991-07-01

    Strategic planning needs to be done as one of the integral steps in fulfilling our overall Departmental mission. The role of strategic planning is to assure that the longer term destinations, goals, and objectives which the programs and activities of the Department are striving towards are the best we can envision today so that our courses can then be set to move in those directions. Strategic planning will assist the Secretary, Deputy Secretary, and Under Secretary in setting the long-term directions and policies for the Department and in making final decisions on near-term priorities and resource allocations. It will assist program developers and implementors by providing the necessary guidance for multi-year program plans and budgets. It is one of the essential steps in the secretary's Strategic Planning Initiative. The operational planning most of us are so familiar with deals with how to get things done and with the resources needed (people, money, facilities, time) to carry out tasks. Operating plans like budgets, capital line item projects, R D budgets, project proposals, etc., are vital to the mission of the Department. They deal, however, with how to carry out programs to achieve some objective or budget assumption. Strategic planning deals with the prior question of what it is that should be attempted. It deals with what objectives the many programs and activities of the Department of Department should be striving toward. The purpose of this document is to provide guidance to those organizations and personnel starting the process for the first time as well as those who have prepared strategic plans in the past and now wish to review and update them. This guideline should not be constructed as a rigid, restrictive or confining rulebook. Each organization is encouraged to develop such enhancements as they think may be useful in their planning. The steps outlined in this document represent a very simplified approach to strategic planning. 9 refs.

  13. Strategic forces briefing

    SciTech Connect

    Bing, G.; Chrzanowski, P.; May, M.; Nordyke, M.

    1989-04-06

    The Strategic Forces Briefing'' is our attempt, accomplished over the past several months, to outline and highlight the more significant strategic force issues that must be addressed in the near future. Some issues are recurrent: the need for an effective modernized Triad and a constant concern for force survivability. Some issues derive from arms control: the Strategic Arms Reduction Talks (SALT) are sufficiently advanced to set broad numerical limits on forces, but not so constraining as to preclude choices among weapon systems and deployment modes. Finally, a new administration faced with serious budgetary problems must strive for the most effective strategic forces limited dollars can buy and support. A review of strategic forces logically begins with consideration of the missions the forces are charged with. We begin the briefing with a short review of targeting policy and implementation within the constraints of available unclassified information. We then review each element of the Triad with sections on SLBMs, ICBMs, and Air-Breathing (bomber and cruise missile) systems. A short section at the end deals with the potential impact of strategic defense on offensive force planning. We consider ABM, ASAT, and air defense; but we do not attempt to address the technical issues of strategic defense per se. The final section gives a brief overview of the tritium supply problem. We conclude with a summary of recommendations that emerge from our review. The results of calculation on the effectiveness of various weapon systems as a function of cost that are presented in the briefing are by Paul Chrzanowski.

  14. lck-Driven Cre Expression Alters T Cell Development in the Thymus and the Frequencies and Functions of Peripheral T Cell Subsets.

    PubMed

    Carow, Berit; Gao, Yu; Coquet, Jonathan; Reilly, Marie; Rottenberg, Martin E

    2016-09-15

    Conditional gene targeting using the bacteriophage-derived Cre recombinase is widely applied for functional gene studies in mice. Mice transgenic for Cre under the control of the lck gene promoter are used to study the role of loxP-targeted genes in T cell development and function. In this article, we show a striking 65% reduction in cellularity, preferential development of γδ versus αβ T cells, and increased expression of IL-7R in the thymus of mice expressing Cre under the proximal lck promoter (lck-cre(+) mice). The transition from CD4/CD8 double-negative to double-positive cells was blocked, and lck-cre(+) double-positive cells were more prone to apoptosis and showed higher levels of Cre expression. Importantly, numbers of naive T cells were reduced in spleens and lymph nodes of lck-cre(+) mice. In contrast, frequencies of γδ T cells, CD44(+)CD62L(-) effector T cells, and Foxp3(+) regulatory T cells were elevated, as was the frequency of IFN-γ-secreting CD4(+) and CD8(+) T cells. A literature survey of 332 articles that used lck-cre(+) mice for deletion of floxed genes indicated that results are statistically influenced by the control used (lck-cre(+) or lck-cre(-)), more frequently resembling the lck-cre(+) phenotype described in this article if lck-cre(-) controls were used. Altogether, care should be taken when interpreting published results and to properly control targeted gene deletions using the lck-cre(+) strain. PMID:27503210

  15. Strategic arms limitation

    SciTech Connect

    Greb, G.A.; Johnson, G.W.

    1983-01-01

    Since 1969, the focus of the Soviet-American arms control process has been on limiting the numbers and sizes of both defensive and offensive strategic systems. The format for this effort has been the Strategic Arms Limitations Talks (SALT) and more recently the Strategic Arms Reduction Talks (START). Both sides came to these negotiations convinced that nuclear arsenals had grown so large that some form of mutual restraint was needed. Although the SALT/START process has been slow and ponderous, it has produced several concrete agreements and collateral benefits. The 1972 ABM Treaty restricts the deployment of ballistic missile defense systems, the 1972 Interim Agreement places a quantitative freeze on each side's land based and sea based strategic launchers, and the as yet unratified 1979 SALT II Treaty sets numerical limits on all offensive strategic systems and sublimits on MIRVed systems. Collateral benefits include improved verification procedures, working definitions and counting rules, and permanent bureaucratic apparatus that enhance stability and increase the chances for achieving additional agreements.

  16. NASA strategic plan

    NASA Technical Reports Server (NTRS)

    1994-01-01

    The NASA Strategic Plan is a living document. It provides far-reaching goals and objectives to create stability for NASA's efforts. The Plan presents NASA's top-level strategy: it articulates what NASA does and for whom; it differentiates between ends and means; it states where NASA is going and what NASA intends to do to get there. This Plan is not a budget document, nor does it present priorities for current or future programs. Rather, it establishes a framework for shaping NASA's activities and developing a balanced set of priorities across the Agency. Such priorities will then be reflected in the NASA budget. The document includes vision, mission, and goals; external environment; conceptual framework; strategic enterprises (Mission to Planet Earth, aeronautics, human exploration and development of space, scientific research, space technology, and synergy); strategic functions (transportation to space, space communications, human resources, and physical resources); values and operating principles; implementing strategy; and senior management team concurrence.

  17. Strategic Analysis of Terrorism

    NASA Astrophysics Data System (ADS)

    Arce, Daniel G.; Sandler, Todd

    Two areas that are increasingly studied in the game-theoretic literature on terrorism and counterterrorism are collective action and asymmetric information. One contribution of this chapter is a survey and extension of continuous policy models with differentiable payoff functions. In this way, policies can be characterized as strategic substitutes (e. g., proactive measures), or strategic complements (e. g., defensive measures). Mixed substitute-complement models are also introduced. We show that the efficiency of counterterror policy depends upon (i) the strategic substitutes-complements characterization, and (ii) who initiates the action. Surprisingly, in mixed-models the dichotomy between individual and collective action may disappear. A second contribution is the consideration of a signaling model where indiscriminant spectacular terrorist attacks may erode terrorists’ support among its constituency, and proactive government responses can create a backlash effect in favor of terrorists. A novel equilibrium of this model reflects the well-documented ineffectiveness of terrorism in achieving its stated goals.

  18. Generation and Characterization of an Nse-CreERT2 Transgenic Line Suitable for Inducible Gene Manipulation in Cerebellar Granule Cells

    PubMed Central

    Pohlkamp, Theresa; Steller, Laura; May, Petra; Günther, Thomas; Schüle, Roland; Frotscher, Michael

    2014-01-01

    We created an Nse-CreERT2 mouse line expressing the tamoxifen-inducible CreERT2 recombinase under the control of the neuron-specific enolase (Nse) promoter. By using Cre reporter lines we could show that this Nse-CreERT2 line has recombination activity in the granule cells of all cerebellar lobules as well as in postmitotic granule cell precursors in the external granular layer of the developing cerebellum. A few hippocampal dentate gyrus granule cells showed Cre-mediated recombination as well. Cre activity could be induced in both the developing and adult mouse brain. The established mouse line constitutes a valuable tool to study the function of genes expressed by cerebellar granule cells in the developing and adult brain. In combination with reporter lines it is a useful model to analyze the development and maintenance of the cerebellar architecture including granule cell distribution, migration, and the extension of granule cell fibers in vivo. PMID:24950299

  19. Strategic avionics technology planning

    NASA Technical Reports Server (NTRS)

    Cox, Kenneth J.; Brown, Don C.

    1991-01-01

    NASA experience in development and insertion of technology into programs had led to a recognition that a Strategic Plan for Avionics is needed for space. In the fall of 1989 an Avionics Technology Symposium was held in Williamsburg, Virginia. In early 1990, as a followon, a NASA wide Strategic Avionics Technology Working Group was chartered by NASA Headquarters. This paper will describe the objectives of this working group, technology bridging, and approaches to incentivize both the federal and commercial sectors to move toward rapidly developed, simple, and reliable systems with low life cycle cost.

  20. On operator strategic behavior

    NASA Technical Reports Server (NTRS)

    Hancock, P. A.

    1991-01-01

    Deeper and more detailed knowledge as to how human operators such as pilots respond, singly and in groups, to demands on their performance which arise from technical systems will support the manipulation of such systems' design in order to accommodate the foibles of human behavior. Efforts to understand how self-autonomy impacts strategic behavior and such related issues as error generation/recognition/correction are still in their infancy. The present treatment offers both general and aviation-specific definitions of strategic behavior as precursors of prospective investigations.

  1. Microgravity strategic planning exercise

    NASA Technical Reports Server (NTRS)

    Halpern, Richard; Downey, Jim; Harvey, Harold

    1991-01-01

    The Center for Space and Advanced Technology supported a planning exercise for the Microgravity Program management at the Marshall Space Flight Center. The effort focused on the status of microgravity work at MSFC and elsewhere with the objective of preparing a goal-oriented strategic planning document which could be used for informational/brochure purposes. The effort entailed numerous interactions and presentations with Field Center programmatic components and Headquarters personnel. Appropriate material was consolidated in a draft format for a MSFC Strategic Plan.

  2. Cloning-free genome engineering in Sinorhizobium meliloti advances applications of Cre/loxP site-specific recombination.

    PubMed

    Döhlemann, Johannes; Brennecke, Meike; Becker, Anke

    2016-09-10

    The soil-dwelling α-proteobacterium Sinorhizobium meliloti serves as model for studies of symbiotic nitrogen fixation, a highly important process in sustainable agriculture. Here, we report advancements of the genetic toolbox accelerating genome editing in S. meliloti. The hsdMSR operon encodes a type-I restriction-modification (R-M) system. Transformation of S. meliloti is counteracted by the restriction endonuclease HsdR degrading DNA which lacks the appropriate methylation pattern. We provide a stable S. meliloti hsdR deletion mutant showing enhanced transformation with Escherichia coli-derived plasmid DNA and demonstrate that using an E. coli plasmid donor, expressing S. meliloti methyl transferase genes, is an alternative strategy of increasing the transformation efficiency of S. meliloti. Furthermore, we devise a novel cloning-free genome editing (CFGE) method for S. meliloti, Agrobacterium tumefaciens and Xanthomonas campestris, and demonstrate the applicability of this method for intricate applications of the Cre/lox recombination system in S. meliloti. An enhanced Cre/lox system, allowing for serial deletions of large genomic regions, was established. An assay of lox spacer mutants identified a set of lox sites mediating specific recombination. The availability of several non-promiscuous Cre recognition sites enables simultaneous specific Cre/lox recombination events. CFGE combined with Cre/lox recombination is put forward as powerful approach for targeted genome editing, involving serial steps of manipulation to expedite the genetic accessibility of S. meliloti as chassis. PMID:27393468

  3. Use of Cre/loxP recombination to swap cell binding motifs on the adenoviral capsid protein IX

    SciTech Connect

    Poulin, Kathy L.; Tong, Grace; Vorobyova, Olga; Pool, Madeline; Kothary, Rashmi; Parks, Robin J.

    2011-11-25

    We used Cre/loxP recombination to swap targeting ligands present on the adenoviral capsid protein IX (pIX). A loxP-flanked sequence encoding poly-lysine (pK-binds heparan sulfate proteoglycans) was engineered onto the 3'-terminus of pIX, and the resulting fusion protein allowed for routine virus propagation. Growth of this virus on Cre-expressing cells removed the pK coding sequence, generating virus that could only infect through alternative ligands, such as a tyrosine kinase receptor A (TrkA)-binding motif engineered into the capsid fibre protein for enhanced infection of neuronal cells. We used a similar approach to swap the pK motif on pIX for a sequence encoding a single-domain antibody directed towards CD66c for targeted infection of cancer cells; Cre-mediated removal of the pK-coding sequence simultaneously placed the single-domain antibody coding sequence in frame with pIX. Thus, we have developed a simple method to propagate virus lacking native viral tropism but containing cell-specific binding ligands. - Highlights: > We describe a method to grow virus lacking native tropism but containing novel cell-binding ligands. > Cre/loxP recombination was used to modify the adenovirus genome. > A targeting ligand present on capsid protein IX was removed or replaced using recombination. > Cre-loxP was also used to 'swap' the identity of the targeting ligand present on pIX.

  4. Generating conditional mutants to analyze ciliary functions: the use of Cre-lox technology to disrupt cilia in specific organs.

    PubMed

    O'Connor, Amber K; Kesterson, Robert A; Yoder, Bradley K

    2009-01-01

    The list of human disordered associated with cilia dysfunction, the ciliopathies, continues to highlight the importance of understanding the many roles of the long overlooked primary cilium. Much of the insights into the clinical importance of the cilium have come from analyses in model organisms, especially the mouse. However, the early embryonic lethality and severe developmental defects associated with cilia disruption has hindered progress in exploring cilia functions in late development or in adult tissues. This hurdle is being surmounted through the use of conditional alleles of genes encoding ciliary proteins and Cre deletor lines with inducible Cre activity or with lines expressing Cre in a cell-type-specific manner. Results from these approaches are providing important insights into the diverse array of cellular and tissue activities regulated by the cilium. Here we provide a recent account of the Cre/lox strategy. The generation and use of well-designed conditional alleles, as well as careful manipulation of embryonic stem cells are discussed. We also provide specific examples to illustrate the use of Cre/lox approaches to evaluate ciliary function in several tissues. With the recent characterization of multiple cilia proteomes along with efforts of several consortia to generate conditional alleles of all genes in the mouse, further use of conditional mutation approaches promise to yield many advances and surprises as we explore the functions of this increasingly complex organelle. PMID:20409823

  5. Flavonoids and Auxin Transport Inhibitors Rescue Symbiotic Nodulation in the Medicago truncatula Cytokinin Perception Mutant cre1

    PubMed Central

    Ng, Jason Liang Pin; Hassan, Samira; Truong, Thy T.; Hocart, Charles H.; Laffont, Carole; Frugier, Florian; Mathesius, Ulrike

    2015-01-01

    Initiation of symbiotic nodules in legumes requires cytokinin signaling, but its mechanism of action is largely unknown. Here, we tested whether the failure to initiate nodules in the Medicago truncatula cytokinin perception mutant cre1 (cytokinin response1) is due to its altered ability to regulate auxin transport, auxin accumulation, and induction of flavonoids. We found that in the cre1 mutant, symbiotic rhizobia cannot locally alter acro- and basipetal auxin transport during nodule initiation and that these mutants show reduced auxin (indole-3-acetic acid) accumulation and auxin responses compared with the wild type. Quantification of flavonoids, which can act as endogenous auxin transport inhibitors, showed a deficiency in the induction of free naringenin, isoliquiritigenin, quercetin, and hesperetin in cre1 roots compared with wild-type roots 24 h after inoculation with rhizobia. Coinoculation of roots with rhizobia and the flavonoids naringenin, isoliquiritigenin, and kaempferol, or with the synthetic auxin transport inhibitor 2,3,5,-triiodobenzoic acid, rescued nodulation efficiency in cre1 mutants and allowed auxin transport control in response to rhizobia. Our results suggest that CRE1-dependent cytokinin signaling leads to nodule initiation through the regulation of flavonoid accumulation required for local alteration of polar auxin transport and subsequent auxin accumulation in cortical cells during the early stages of nodulation. PMID:26253705

  6. Nerve growth factor enhances the CRE-dependent transcriptional activity activated by nobiletin in PC12 cells.

    PubMed

    Takito, Jiro; Kimura, Junko; Kajima, Koji; Uozumi, Nobuyuki; Watanabe, Makoto; Yokosuka, Akihito; Mimaki, Yoshihiro; Nakamura, Masanori; Ohizumi, Yasushi

    2016-07-01

    Prevention and treatment of Alzheimer disease are urgent problems for elderly people in developed countries. We previously reported that nobiletin, a poly-methoxylated flavone from the citrus peel, improved the symptoms in various types of animal models of memory loss and activated the cAMP responsive element (CRE)-dependent transcription in PC12 cells. Nobiletin activated the cAMP/PKA/MEK/Erk/MAPK signaling pathway without using the TrkA signaling activated by nerve growth factor (NGF). Here, we examined the effect of combination of nobiletin and NGF on the CRE-dependent transcription in PC12 cells. Although NGF alone had little effect on the CRE-dependent transcription, NGF markedly enhanced the CRE-dependent transcription induced by nobiletin. The NGF-induced enhancement was neutralized by a TrkA antagonist, K252a. This effect of NGF was effective on the early signaling event elicited by nobiletin. These results suggested that there was crosstalk between NGF and nobiletin signaling in activating the CRE-dependent transcription in PC12 cells. PMID:27128150

  7. Strategic plan for infrastructure optimization

    SciTech Connect

    Donley, C.D.

    1998-05-27

    This document represents Fluor Daniel Hanford`s and DynCorp`s Tri-Cities Strategic Plan for Fiscal Years 1998--2002, the road map that will guide them into the next century and their sixth year of providing safe and cost effective infrastructure services and support to the Department of Energy (DOE) and the Hanford Site. The Plan responds directly to the issues raised in the FDH/DOE Critical Self Assessment specifically: (1) a strategy in place to give DOE the management (systems) and physical infrastructure for the future; (2) dealing with the barriers that exist to making change; and (3) a plan to right-size the infrastructure and services, and reduce the cost of providing services. The Plan incorporates initiatives from several studies conducted in Fiscal Year 1997 to include: the Systems Functional Analysis, 200 Area Water Commercial Practices Plan, $ million Originated Cost Budget Achievement Plan, the 1OO Area Vacate Plan, the Railroad Shutdown Plan, as well as recommendations from the recently completed Review of Hanford Electrical Utility. These and other initiatives identified over the next five years will result in significant improvements in efficiency, allowing a greater portion of the infrastructure budget to be applied to Site cleanup. The Plan outlines a planning and management process that defines infrastructure services and structure by linking site technical base line data and customer requirements to work scope and resources. The Plan also provides a vision of where Site infrastructure is going and specific initiatives to get there.

  8. Comparison of Three Cre-LoxP Based Paired-End Library Construction Methods

    SciTech Connect

    Peng, Ze; Nath, Nandita; Tritt, Andrew; Liang, Shoudan; Han, James; Pennacchio, Len; Chen, Feng

    2013-03-26

    Paired-end library sequencing has been proven useful in scaffold construction during de novo whole genome shotgun assembly. The ability of generating mate pairs with > 8 Kb insert sizes is especially important for genomes containing long repeats. To make mate paired libraries for next generation sequencing, DNA fragments need to be circularized to bring the ends together. There are several methods that can be used for DNA circulation, namely ligation, hybridization and Cre-LoxP recombination. With higher circularization efficiency with large insert DNA fragments, Cre-LoxP recombination method generally has been used for constructing >8 kb insert size paired-end libraries. Second fragmentation step is also crucial for maintaining high library complexity and uniform genome coverage. Here we will describe the following three fragmentation methods: restriction enzyme digestion, random shearing and nick translation. We will present the comparison results for these three methods. Our data showed that all three methods are able to generate paired-end libraries with greater than 20 kb insert. Advantages and disadvantages of these three methods will be discussed as well.

  9. The Twannberg (Switzerland) IIG iron meteorites: Mineralogy, chemistry, and CRE ages

    NASA Astrophysics Data System (ADS)

    Hofmann, B. A.; Lorenzetti, S.; Eugster, O.; Krähenbühl, U.; Herzog, G.; Serefiddin, F.; Gnos, E.; Eggimann, M.; Wasson, J. T.

    2009-03-01

    The original mass (15915 g) of the Twannberg IIG (low Ni-, high P) iron was found in 1984. Five additional masses (12 to 2488 g) were recovered between 2000 and 2007 in the area. The different masses show identical mineralogy consisting of kamacite single crystals with inclusions of three types of schreibersite crystals: cm-sized skeletal (10.5% Ni), lamellar (17.2% Ni), and 1-3 × 10 µm-sized microprismatic (23.9% Ni). Masses I and II were compared in detail and have virtually identical microstructure, hardness, chemical composition, cosmic-ray exposure (CRE) ages, and 10Be and 26Al activities. Bulk concentrations of 5.2% Ni and 2.0% P were calculated. The preatmospheric mass is estimated to have been at least 11,000 kg. The average CRE age for the different Twannberg samples is 230 ± 50 Ma. Detrital terrestrial mineral grains in the oxide rinds of the three larger masses indicate that they oxidized while they were incorporated in a glacial till deposited by the RhÙne glacier during the last glaciation (Würm). The find location of mass I is located at the limit of glaciation where the meteorite may have deposited after transport by the glacier over considerable distance. All evidence indicates pairing of the six masses, which may be part of a larger shower as is indicated by the large inferred pre-atmospheric mass.

  10. A Cre-dependent GCaMP3 reporter mouse for neuronal imaging in vivo

    PubMed Central

    Zariwala, Hatim A.; Borghuis, Bart G.; Hoogland, Tycho M.; Madisen, Linda; Tian, Lin; De Zeeuw, Chris I.; Zeng, Hongkui; Looger, Loren L.; Svoboda, Karel; Chen, Tsai-Wen

    2012-01-01

    Fluorescent calcium indicator proteins, such as GCaMP3, allow imaging of activity in genetically defined neuronal populations. GCaMP3 can be expressed using various gene delivery methods, such as viral infection or electroporation. However, these methods are invasive and provide inhomogeneous and non-stationary expression. Here we developed a genetic reporter mouse, Ai38, which expresses GCaMP3 in a Cre-dependent manner from the ROSA26 locus, driven by a strong CAG promoter. Crossing Ai38 with appropriate Cre mice produced robust GCaMP3 expression in defined cell populations in the retina, cortex and cerebellum. In the primary visual cortex, visually-evoked GCaMP3 signals showed normal orientation and direction selectivity. GCaMP3 signals were rapid, compared to virally expressed GCaMP3 and synthetic calcium indicators. In the retina, Ai38 allowed imaging spontaneous calcium waves in starburst amacrine cells during development, and light-evoked responses in ganglion cells in adult tissue. Our results show that the Ai38 reporter mouse provides a flexible method for targeted expression of GCaMP3. PMID:22378886

  11. Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain.

    PubMed

    Deverman, Benjamin E; Pravdo, Piers L; Simpson, Bryan P; Kumar, Sripriya Ravindra; Chan, Ken Y; Banerjee, Abhik; Wu, Wei-Li; Yang, Bin; Huber, Nina; Pasca, Sergiu P; Gradinaru, Viviana

    2016-02-01

    Recombinant adeno-associated viruses (rAAVs) are commonly used vehicles for in vivo gene transfer. However, the tropism repertoire of naturally occurring AAVs is limited, prompting a search for novel AAV capsids with desired characteristics. Here we describe a capsid selection method, called Cre recombination-based AAV targeted evolution (CREATE), that enables the development of AAV capsids that more efficiently transduce defined Cre-expressing cell populations in vivo. We use CREATE to generate AAV variants that efficiently and widely transduce the adult mouse central nervous system (CNS) after intravenous injection. One variant, AAV-PHP.B, transfers genes throughout the CNS with an efficiency that is at least 40-fold greater than that of the current standard, AAV9 (refs. 14,15,16,17), and transduces the majority of astrocytes and neurons across multiple CNS regions. In vitro, it transduces human neurons and astrocytes more efficiently than does AAV9, demonstrating the potential of CREATE to produce customized AAV vectors for biomedical applications. PMID:26829320

  12. BioCreAtIvE Task 1A: gene mention finding evaluation

    PubMed Central

    Yeh, Alexander; Morgan, Alexander; Colosimo, Marc; Hirschman, Lynette

    2005-01-01

    Background The biological research literature is a major repository of knowledge. As the amount of literature increases, it will get harder to find the information of interest on a particular topic. There has been an increasing amount of work on text mining this literature, but comparing this work is hard because of a lack of standards for making comparisons. To address this, we worked with colleagues at the Protein Design Group, CNB-CSIC, Madrid to develop BioCreAtIvE (Critical Assessment for Information Extraction in Biology), an open common evaluation of systems on a number of biological text mining tasks. We report here on task 1A, which deals with finding mentions of genes and related entities in text. "Finding mentions" is a basic task, which can be used as a building block for other text mining tasks. The task makes use of data and evaluation software provided by the (US) National Center for Biotechnology Information (NCBI). Results 15 teams took part in task 1A. A number of teams achieved scores over 80% F-measure (balanced precision and recall). The teams that tried to use their task 1A systems to help on other BioCreAtIvE tasks reported mixed results. Conclusion The 80% plus F-measure results are good, but still somewhat lag the best scores achieved in some other domains such as newswire, due in part to the complexity and length of gene names, compared to person or organization names in newswire. PMID:15960832

  13. Nuclear DISC1 regulates CRE-mediated gene transcription and sleep homeostasis in the fruit fly.

    PubMed

    Sawamura, N; Ando, T; Maruyama, Y; Fujimuro, M; Mochizuki, H; Honjo, K; Shimoda, M; Toda, H; Sawamura-Yamamoto, T; Makuch, L A; Hayashi, A; Ishizuka, K; Cascella, N G; Kamiya, A; Ishida, N; Tomoda, T; Hai, T; Furukubo-Tokunaga, K; Sawa, A

    2008-12-01

    Disrupted-in-schizophrenia-1 (DISC1) is one of major susceptibility factors for a wide range of mental illnesses, including schizophrenia, bipolar disorder, major depression and autism spectrum conditions. DISC1 is located in several subcellular domains, such as the centrosome and the nucleus, and interacts with various proteins, including NudE-like (NUDEL/NDEL1) and activating transcription factor 4 (ATF4)/CREB2. Nevertheless, a role for DISC1 in vivo remains to be elucidated. Therefore, we have generated a Drosophila model for examining normal functions of DISC1 in living organisms. DISC1 transgenic flies with preferential accumulation of exogenous human DISC1 in the nucleus display disturbance in sleep homeostasis, which has been reportedly associated with CREB signaling/CRE-mediated gene transcription. Thus, in mammalian cells, we characterized nuclear DISC1, and identified a subset of nuclear DISC1 that colocalizes with the promyelocytic leukemia (PML) bodies, a nuclear compartment for gene transcription. Furthermore, we identified three functional cis-elements that regulate the nuclear localization of DISC1. We also report that DISC1 interacts with ATF4/CREB2 and a corepressor N-CoR, modulating CRE-mediated gene transcription. PMID:18762802

  14. BAC modification through serial or simultaneous use of CRE/Lox technology.

    PubMed

    Parrish, Mark; Unruh, Jay; Krumlauf, Robb

    2011-01-01

    Bacterial Artificial Chromosomes (BACs) are vital tools in mouse genomic analyses because of their ability to propagate large inserts. The size of these constructs, however, prevents the use of conventional molecular biology techniques for modification and manipulation. Techniques such as recombineering and Cre/Lox methodologies have thus become heavily relied upon for such purposes. In this work, we investigate the applicability of Lox variant sites for serial and/or simultaneous manipulations of BACs. We show that Lox spacer mutants are very specific, and inverted repeat variants reduce Lox reaction rates through reducing the affinity of Cre for the site, while retaining some functionality. Employing these methods, we produced serial modifications encompassing four independent changes which generated a mouse HoxB BAC with fluorescent reporter proteins inserted into four adjacent Hox genes. We also generated specific, simultaneous deletions using combinations of spacer variants and inverted repeat variants. These techniques will facilitate BAC manipulations and open a new repertoire of methods for BAC and genome manipulation. PMID:21197414

  15. Generalized bacterial genome editing using mobile group II introns and Cre-lox

    PubMed Central

    Enyeart, Peter J; Chirieleison, Steven M; Dao, Mai N; Perutka, Jiri; Quandt, Erik M; Yao, Jun; Whitt, Jacob T; Keatinge-Clay, Adrian T; Lambowitz, Alan M; Ellington, Andrew D

    2013-01-01

    Efficient bacterial genetic engineering approaches with broad-host applicability are rare. We combine two systems, mobile group II introns (‘targetrons') and Cre/lox, which function efficiently in many different organisms, into a versatile platform we call GETR (Genome Editing via Targetrons and Recombinases). The introns deliver lox sites to specific genomic loci, enabling genomic manipulations. Efficiency is enhanced by adding flexibility to the RNA hairpins formed by the lox sites. We use the system for insertions, deletions, inversions, and one-step cut-and-paste operations. We demonstrate insertion of a 12-kb polyketide synthase operon into the lacZ gene of Escherichia coli, multiple simultaneous and sequential deletions of up to 120 kb in E. coli and Staphylococcus aureus, inversions of up to 1.2 Mb in E. coli and Bacillus subtilis, and one-step cut-and-pastes for translocating 120 kb of genomic sequence to a site 1.5 Mb away. We also demonstrate the simultaneous delivery of lox sites into multiple loci in the Shewanella oneidensis genome. No selectable markers need to be placed in the genome, and the efficiency of Cre-mediated manipulations typically approaches 100%. PMID:24002656

  16. Optimized second-generation CRY2-CIB dimerizers and photoactivatable Cre recombinase.

    PubMed

    Taslimi, Amir; Zoltowski, Brian; Miranda, Jose G; Pathak, Gopal P; Hughes, Robert M; Tucker, Chandra L

    2016-06-01

    Arabidopsis thaliana cryptochrome 2 (AtCRY2), a light-sensitive photosensory protein, was previously adapted for use in controlling protein-protein interactions through light-dependent binding to a partner protein, CIB1. While the existing CRY2-CIB dimerization system has been used extensively for optogenetic applications, some limitations exist. Here, we set out to optimize function of the CRY2-CIB system by identifying versions of CRY2-CIB that are smaller, show reduced dark interaction, and maintain longer or shorter signaling states in response to a pulse of light. We describe minimal functional CRY2 and CIB1 domains maintaining light-dependent interaction and new signaling mutations affecting AtCRY2 photocycle kinetics. The latter work implicates an α13-α14 turn motif within plant CRYs whose perturbation alters signaling-state lifetime. Using a long-lived L348F photocycle mutant, we engineered a second-generation photoactivatable Cre recombinase, PA-Cre2.0, that shows five-fold improved dynamic range, allowing robust recombination following exposure to a single, brief pulse of light. PMID:27065233

  17. A Cre-dependent GCaMP3 reporter mouse for neuronal imaging in vivo.

    PubMed

    Zariwala, Hatim A; Borghuis, Bart G; Hoogland, Tycho M; Madisen, Linda; Tian, Lin; De Zeeuw, Chris I; Zeng, Hongkui; Looger, Loren L; Svoboda, Karel; Chen, Tsai-Wen

    2012-02-29

    Fluorescent calcium indicator proteins, such as GCaMP3, allow imaging of activity in genetically defined neuronal populations. GCaMP3 can be expressed using various gene delivery methods, such as viral infection or electroporation. However, these methods are invasive and provide inhomogeneous and nonstationary expression. Here, we developed a genetic reporter mouse, Ai38, which expresses GCaMP3 in a Cre-dependent manner from the ROSA26 locus, driven by a strong CAG promoter. Crossing Ai38 with appropriate Cre mice produced robust GCaMP3 expression in defined cell populations in the retina, cortex, and cerebellum. In the primary visual cortex, visually evoked GCaMP3 signals showed normal orientation and direction selectivity. GCaMP3 signals were rapid, compared with virally expressed GCaMP3 and synthetic calcium indicators. In the retina, Ai38 allowed imaging spontaneous calcium waves in starburst amacrine cells during development, and light-evoked responses in ganglion cells in adult tissue. Our results show that the Ai38 reporter mouse provides a flexible method for targeted expression of GCaMP3. PMID:22378886

  18. Cre-inducible human CD59 mediates rapid cell ablation after intermedilysin administration.

    PubMed

    Feng, Dechun; Dai, Shen; Liu, Fengming; Ohtake, Yosuke; Zhou, Zhou; Wang, Hua; Zhang, Yonggang; Kearns, Alison; Peng, Xiao; Zhu, Faliang; Hayat, Umar; Li, Man; He, Yong; Xu, Mingjiang; Zhao, Chunling; Cheng, Min; Zhang, Lining; Wang, Hong; Yang, Xiaofeng; Ju, Cynthia; Bryda, Elizabeth C; Gordon, Jennifer; Khalili, Kamel; Hu, Wenhui; Li, Shuxin; Qin, Xuebin; Gao, Bin

    2016-06-01

    Cell ablation is a powerful tool for studying cell lineage and/or function; however, current cell-ablation models have limitations. Intermedilysin (ILY), a cytolytic pore-forming toxin that is secreted by Streptococcus intermedius, lyses human cells exclusively by binding to the human complement regulator CD59 (hCD59), but does not react with CD59 from nonprimates. Here, we took advantage of this feature of ILY and developed a model of conditional and targeted cell ablation by generating floxed STOP-CD59 knockin mice (ihCD59), in which expression of human CD59 only occurs after Cre-mediated recombination. The administration of ILY to ihCD59+ mice crossed with various Cre-driver lines resulted in the rapid and specific ablation of immune, epithelial, or neural cells without off-target effects. ILY had a large pharmacological window, which allowed us to perform dose-dependent studies. Finally, the ILY/ihCD59-mediated cell-ablation method was tested in several disease models to study immune cell functionalities, hepatocyte and/or biliary epithelial damage and regeneration, and neural cell damage. Together, the results of this study demonstrate the utility of the ihCD59 mouse model for studying the effects of cell ablation in specific organ systems in a variety of developmental and disease states. PMID:27159394

  19. 28 CFR 902.2 - Raising disputes.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 28 Judicial Administration 2 2014-07-01 2014-07-01 false Raising disputes. 902.2 Section 902.2 Judicial Administration NATIONAL CRIME PREVENTION AND PRIVACY COMPACT COUNCIL DISPUTE ADJUDICATION PROCEDURES § 902.2 Raising disputes. (a) Cognizable disputes may be based upon: (1) A claim that the...

  20. 28 CFR 902.2 - Raising disputes.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 28 Judicial Administration 2 2010-07-01 2010-07-01 false Raising disputes. 902.2 Section 902.2 Judicial Administration NATIONAL CRIME PREVENTION AND PRIVACY COMPACT COUNCIL DISPUTE ADJUDICATION PROCEDURES § 902.2 Raising disputes. (a) Cognizable disputes may be based upon: (1) A claim that the...

  1. 28 CFR 902.2 - Raising disputes.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 28 Judicial Administration 2 2011-07-01 2011-07-01 false Raising disputes. 902.2 Section 902.2 Judicial Administration NATIONAL CRIME PREVENTION AND PRIVACY COMPACT COUNCIL DISPUTE ADJUDICATION PROCEDURES § 902.2 Raising disputes. (a) Cognizable disputes may be based upon: (1) A claim that the...

  2. 28 CFR 902.2 - Raising disputes.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 28 Judicial Administration 2 2013-07-01 2013-07-01 false Raising disputes. 902.2 Section 902.2 Judicial Administration NATIONAL CRIME PREVENTION AND PRIVACY COMPACT COUNCIL DISPUTE ADJUDICATION PROCEDURES § 902.2 Raising disputes. (a) Cognizable disputes may be based upon: (1) A claim that the...

  3. 28 CFR 902.2 - Raising disputes.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 28 Judicial Administration 2 2012-07-01 2012-07-01 false Raising disputes. 902.2 Section 902.2 Judicial Administration NATIONAL CRIME PREVENTION AND PRIVACY COMPACT COUNCIL DISPUTE ADJUDICATION PROCEDURES § 902.2 Raising disputes. (a) Cognizable disputes may be based upon: (1) A claim that the...

  4. Children's Difficulty with Raising: A Performance Account

    ERIC Educational Resources Information Center

    Choe, Jinsun; Deen, Kamil

    2016-01-01

    This article explores English-speaking children's acquisition of raising structures with an experiencer (e.g.," John seems to Mary to be happy"). We review and address previously unnoticed issues in the methodologies of existing studies testing the acquisition of raising, thus providing a more reliable picture of children's abilities…

  5. Raising Sensitive Issues in a Team

    ERIC Educational Resources Information Center

    Lindoerfer, Dennis

    2008-01-01

    Have you ever wondered how to deal with a sensitive issue within your team? For example, how do you raise the issue that the women rarely get listened to? How do you bring up your observation that the team members from Marketing always dominate the meetings? This guidebook focuses on ways to determine whether to raise such an issue in a team…

  6. Strategic Enrollment Management Revolution.

    ERIC Educational Resources Information Center

    Black, Jim, Ed.

    The chapters of this collection describe a transformational period in higher education. The paradigm shift in higher education requires new organizational models, new strategies, new resources, and new tools, with new ways of thinking, especially about strategic enrollment management (SEM). The chapters of part 1, "The Contextual Framework for…

  7. The Strategic Revolution.

    PubMed

    Gardner, Andy

    2016-09-01

    On the 40(th) anniversary of the publication of Richard Dawkins's The Selfish Gene, we explore the origins of cynical, strategic thinking in evolutionary biology, investigate how this illuminated the sexual and social lives of animals, and assess Dawkins's suggestion that evolution is best understood by taking the gene's-eye view. PMID:27610556

  8. Strategic BI for All

    ERIC Educational Resources Information Center

    Raths, David

    2008-01-01

    Implementing a complex business intelligence (BI) system at a small school or one with limited resources can seem daunting. For small to midsize schools and community colleges, a strategic BI initiative may still be an elusive goal. This article discusses how schools with limited resources are making the dream a reality.

  9. Strategic Human Resource Development.

    ERIC Educational Resources Information Center

    Garavan, Thomas N.

    1991-01-01

    Reviews literature on strategic human resource development (HRD) focusing on the characteristics of such activities, conditions necessary for the promotion of HRD, and the benefits to an organization pursuing such activities. Empirical evidence is presented on HRD policy formation and planning processes in Irish high technology companies. (JOW)

  10. Strategic Education Research Partnership.

    ERIC Educational Resources Information Center

    Donovan, M. S., Ed.; Wigdor, A. K., Ed.; Snow, C. E., Ed.

    This book is a proposal for the Strategic Education Research Partnership (SERP), a large-scale coherent program of research and development carried out through a partnership between researchers and practitioners. The program would put the problems of educational practice at its center and focus on carrying research and development through all the…

  11. Using Collaborative Strategic Reading.

    ERIC Educational Resources Information Center

    Klingner, Janette K.; Vaughn, Sharon

    1998-01-01

    Describes collaborative strategic reading (CSR), a technique for teaching students, such as those with learning disabilities, reading comprehension and vocabulary skills in a cooperative setting. Covers teaching the four strategies of CSR (preview, click and clunk, get the gist, and wrap up), as well as teaching students cooperative learning group…

  12. Strategic Marketing for Agribusiness.

    ERIC Educational Resources Information Center

    Welch, Mary A., Ed.

    1993-01-01

    The steps for strategic market planning are discussed including: (1) assessing the situation with market conditions, customers, competitors, and your firm; and (2) crafting a strategy to prioritize target markets, develop a core strategy, and create a marketing mix. Examples of agribusiness successes are presented. The booklet concludes with a…

  13. ACNW - 1998 strategic plan

    SciTech Connect

    1998-04-01

    This plan provides strategic direction to The Advisory Committee on Nuclear Waste (ACNW) in 1998 and beyond for focusing on issues most important to the NRC in carrying out its mission of protecting public health and safety, promoting the common defense and security, and protecting the environment.

  14. EMSL Strategic Plan 2008

    SciTech Connect

    Campbell, Allison A.

    2008-08-15

    This Strategic Plan is EMSL’s template for achieving our vision of simultaneous excellence in all aspects of our mission as a national scientific user facility. It reflects our understanding of the long-term stewardship we must work toward to meet the scientific challenges of the Department of Energy and the nation. During the next decade, we will implement the strategies contained in this Plan, working closely with the scientific community, our advisory committees, DOE’s Office of Biological and Environmental Research, and other key stakeholders. This Strategic Plan is fully aligned with the strategic plans of DOE and its Office of Science. We recognize that shifts in science and technology, national priorities, and resources made available through the Federal budget process create planning uncertainties and, ultimately, a highly dynamic planning environment. Accordingly, this Strategic Plan should be viewed as a living document for which we will continually evaluate changing needs and opportunities posed by our stakeholders (i.e., DOE, users, staff, advisory committees), work closely with them to understand and respond to those changes, and align our strategy accordingly.

  15. Strategic Tutor Monitoring.

    ERIC Educational Resources Information Center

    Chee-kwong, Kenneth Chao

    1996-01-01

    Discusses effective tutor monitoring strategies based on experiences at the Open Learning Institute of Hong Kong. Highlights include key performance and strategic control points; situational factors, including tutor expectations and relevant culture; Theory X versus Theory Y leadership theories; and monitoring relationships with tutors. (LRW)

  16. External Strategic Planning Conference.

    ERIC Educational Resources Information Center

    Los Angeles Community Coll. District, CA. Office of Research and Planning.

    In response to a community reputation that has grown increasingly negative, the Los Angeles Community College District (LACCD) devised a strategic plan in 1998 to improve its programs and services and assure the educational success of its students. The planning process involved several steps: (1) revisiting the district mission statement; (2)…

  17. Manufacturing research strategic plan

    SciTech Connect

    1995-11-01

    This plan provides an overall strategic roadmap for the DOE-defense programs advanced manufacturing research program which supports the national science based stockpile stewardship program. This plan represents a vision required to develop the knowledge base needed to ensure an enduring national capability to rapidly and effectively manufacture nuclear weapons.

  18. Grappling with Strategic Dissonance.

    ERIC Educational Resources Information Center

    Dowie, Sandra

    2002-01-01

    Presents a case study of the Virtual Retina project (an instructional CD-ROM for ophthalmology students) at the University of Alberta as an example of strategic dissonance in an educational technology unit. Offers methods to analyze the external competitive environment and internal capabilities of educational technology units. (EV)

  19. The Strategic Assessment Model.

    ERIC Educational Resources Information Center

    Glazner, Steve, Ed.

    This book presents six papers focusing on the application of the strategic assessment model (SAM) to the management of higher education facilities. The papers are part of an ongoing effort by the Association of Higher Education Facilities Officers to provide comparative cost and staffing information and to develop a benchmarking process. The…

  20. Strategic Goals for 2000.

    ERIC Educational Resources Information Center

    Hopper, Toni

    This is the report of the 2000 New Mexico Association of Community Colleges (NMACC) Strategic Planning Conference. Participants worked in small groups to identify the most probable and the most desirable world for the year 2000. Results for the most probable world include: (1) education would be available to many consumers, using many delivery…

  1. Strategic Marketing for Presidents.

    ERIC Educational Resources Information Center

    Pappas, Richard J., Ed.

    Designed to inform the marketing efforts of community college presidents, this document describes the importance of marketing, presents a targeted approach, and outlines the specific roles and skills needed by the president to ensure successful efforts and effective institutions. The first chapter, "Developing a Marketing-Strategic Plan," by…

  2. Being Strategic in HE Management

    ERIC Educational Resources Information Center

    West, Andrew

    2008-01-01

    The call to be strategic--and with it the concept of strategic management--can bring to mind a wide range of definitions, and there is now a huge array of academic literature supporting the different schools of thought. At a basic level, however, strategic thinking is probably most simply about focusing on the whole, rather than the part. In…

  3. The Possibilities of Strategic Finance

    ERIC Educational Resources Information Center

    Chaffee, Ellen

    2010-01-01

    Strategic finance is aligning financial decisions--regarding revenues, creating and maintaining institutional assets, and using those assets--with the institution's mission and strategic plan. The concept known as "strategic finance" increasingly is being seen as a useful perspective for helping boards and presidents develop a sustainable…

  4. Strategic Planning and Financial Management

    ERIC Educational Resources Information Center

    Conneely, James F.

    2010-01-01

    Strong financial management is a strategy for strategic planning success in student affairs. It is crucial that student affairs professionals understand the necessity of linking their strategic planning with their financial management processes. An effective strategic planner needs strong financial management skills to implement the plan over…

  5. Strategic Partnerships in Higher Education

    ERIC Educational Resources Information Center

    Ortega, Janet L.

    2013-01-01

    The purpose of this study was to investigate the impacts of strategic partnerships between community colleges and key stakeholders; to specifically examine strategic partnerships; leadership decision-making; criteria to evaluate strategic partnerships that added value to the institution, value to the students, faculty, staff, and the local…

  6. Strategic arms limitation

    NASA Astrophysics Data System (ADS)

    Allen Greb, G.; Johnson, Gerald W.

    1983-10-01

    Following World War II, American scientists and politicians proposed in the Baruch plan a radical solution to the problem of nuclear weapons: to eliminate them forever under the auspices of an international nuclear development authority. The Soviets, who as yet did not possess the bomb, rejected this plan. Another approach suggested by Secretary of War Henry Stimson to negotiate directly with the Soviet Union was not accepted by the American leadership. These initial arms limitation failures both reflected and exacerbated the hostile political relationship of the superpowers in the 1950s and 1960s. Since 1969, the more modest focus of the Soviet-American arms control process has been on limiting the numbers and sizes of both defensive and offensive strategic systems. The format for this effort has been the Strategic Arms Limitatins Talks (Salt) and more recently the Strategic Arms Reduction Talks (START). Both sides came to these negotiations convinced that nuclear arsenals had grown so large that some for of mutual restraint was needed. Although the SALT/START process has been slow and ponderous, it has produced several concrete the agreements and collateral benefits. The 1972 ABM Treaty restricts the deployment of ballistic missile defense systems, the 1972 Interim Agreement places a quantitative freeze on each side's land based and sea based strategic launchers, and the as yet unratified 1979 SALT II Treaty sets numerical limits on all offensive strategic systems and sublimits on MIRVed systems. Collateral benefits include improved verification procedures, working definitions and counting rules, and permanent bureaucratic apparatus which enhance stability and increase the chances for achieving additional agreements.

  7. Tissue-Specific Regulation of Oncogene Expression Using Cre-Inducible ROSA26 Knock-In Transgenic Mice.

    PubMed

    Carofino, Brandi L; Justice, Monica J

    2015-01-01

    Cre-inducible mouse models are often utilized for the spatial and temporal expression of oncogenes. With the wide number of Cre recombinase lines available, inducible transgenesis represents a tractable approach to achieve discrete oncogene expression. Here, we describe a protocol for targeting Cre-inducible genes to the ubiquitously expressed ROSA26 locus. Gene targeting provides several advantages over standard transgenic techniques, including a known site of integration and previously characterized pattern of expression. Historically, an inherent instability of ROSA26 targeting vectors has hampered the efficiency of developing ROSA26 knock-in lines. In this protocol, we provide individual steps for utilizing Gateway recombination for cloning as well as detailed instructions for screening targeted ES cell clones. By following this protocol, one can achieve germline transmission of a ROSA26 knock-in line within several months. PMID:26069083

  8. 50 CFR 14.23 - Live farm-raised fish and farm-raised fish eggs.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Exportation at Designated Ports § 14.23 Live farm-raised fish and farm-raised fish eggs. Live farm-raised fish and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3. Except... eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT...

  9. 50 CFR 14.23 - Live farm-raised fish and farm-raised fish eggs.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Exportation at Designated Ports § 14.23 Live farm-raised fish and farm-raised fish eggs. Live farm-raised fish and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3. Except... eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT...

  10. 50 CFR 14.23 - Live farm-raised fish and farm-raised fish eggs.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... Exportation at Designated Ports § 14.23 Live farm-raised fish and farm-raised fish eggs. Live farm-raised fish and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3. Except... eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT...

  11. 50 CFR 14.23 - Live farm-raised fish and farm-raised fish eggs.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... Exportation at Designated Ports § 14.23 Live farm-raised fish and farm-raised fish eggs. Live farm-raised fish and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3. Except... eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT...

  12. 50 CFR 14.23 - Live farm-raised fish and farm-raised fish eggs.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Exportation at Designated Ports § 14.23 Live farm-raised fish and farm-raised fish eggs. Live farm-raised fish and farm-raised fish eggs meet the definition of “bred in captivity” as stated in 50 CFR 17.3. Except... eggs. 14.23 Section 14.23 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT...

  13. Development of selectable marker free, insect resistant, transgenic mustard (Brassica juncea) plants using Cre/lox mediated recombination

    PubMed Central

    2013-01-01

    Background Antibiotic/ herbicide resistant marker genes have been proven to be very useful in plant transformation for the initial selection of desired transgenic events. However, presence of these genes in the genetically modified crops may render the crop less acceptable to the consumers. Among several different approaches, the effectiveness of Cre/lox mediated recombination strategy for selectable marker gene (SMG) elimination has previously been demonstrated by different groups in several plants including Brassica. In the present study exploiting Cre/lox mediated recombination strategy, attempt has been made for selectable marker gene elimination from Allium sativum leaf agglutinin (ASAL) expressing Brassica plants with hemipteran insect resistant phenotype. Results Allium sativum leaf agglutinin (ASAL) linked with lox flanked hygromycin resistant (hpt) gene was introduced in mustard. Cre recombinase gene cassette was also integrated in separate event. A Cre/lox mediated recombination using crossing strategy was adopted to remove the hpt gene from the subsequent generation of selected hybrid events. Reciprocal crosses were made between T1ASAL-lox-hpt-lox and cre-bar plants. Marker gene elimination was confirmed in the resulting F1 hybrid progenies by PCR analysis, using hpt, cre and ASAL specific primers followed by Southern hybridization. In marker free plants, expression of ASAL was also confirmed by western blotting and ELISA analysis. Retention of functionality of expressed ASAL was investigated by agglutination assay using rabbit erythrocytes. Expressed ASAL was also found to be thermo-sensitive. In planta insect bioassay on F1 hybrid progenies exhibited detrimental effect on the performance of devastating target pest, Lipaphis erysimi. The F1 hybrid hpt negative, ASAL positive plants were allowed to self- fertilize to obtain F2 progeny plants. In some of these plants cre gene was found to be segregated out of the ASAL gene by genetic segregation yielding

  14. Normal mammary development and function in mice with Ift88 deleted in MMTV- and K14-Cre expressing cells

    PubMed Central

    2014-01-01

    Background Primary cilia (PC) are non-motile microtubule based organelles present on almost every cell type and are known to serve as critical organizing centers for several signaling pathways crucial to embryonic and postnatal development. Alterations in the Hh pathway, the most studied signaling pathway regulated by PC, affect mammary gland development as well as maintenance of the stem and progenitor cell populations. Results We developed mouse models with deletion of PC in mammary luminal epithelial, basal epithelial, and stromal cells for evaluation of the function of PC in mammary development via MMTV-Cre, K14-Cre, and Prx1-Cre mediated deletion, respectively. The activity of Cre was confirmed using ROSA26 reporters. Mammary stem and progenitor cells were enriched through growth as mammospheres. Adenovirus-Cre mediated deletion of Ift88 was used to determine a role for PC in this population of cells. Disruption of Ift88 and PC were confirmed in using PCR and immunofluorescent methods. Prx1-Cre; Ift88Del mice demonstrated defects in terminal end buds during puberty. However, these Ift88Del glands exhibited typical terminal end bud formation as well as normal ductal histology when transplanted into wild type hosts, indicating that the phenotype observed was not intrinsic to the mammary gland. Furthermore, no discernable alterations to mammary development were observed in MMTV-Cre- or K14-Cre; Ift88Del lines. These mice were able to feed and support several litters of pups even though wide spread depletion of PC was confirmed. Cells grown in mammosphere culture were enriched for PC containing cells suggesting PC are preferentially expressed on mammary stem and progenitor cells. Deletion of Ift88 in mammary epithelial cells resulted in a significant reduction in the number of primary mammospheres established; however, there was no effect on outgrowth of secondary mammospheres in PC-depleted cells. Conclusions PC regulate systemic factors that can affect mammary

  15. Fund Raising. The Guide to Raising Money from Private Sources. Second Edition, Revised and Enlarged.

    ERIC Educational Resources Information Center

    Broce, Thomas E.

    A basic guide and reference to raising funds from private sources is presented that is designed primarily for professional personnel or the administrator employing fund-raising personnel. The first edition of the guide has also been used as a textbook for university courses in fund-raising. This revised edition includes new materials on: capital…

  16. Linking Faculty Raises to College Resources

    ERIC Educational Resources Information Center

    Miller, John

    2006-01-01

    This article discusses how one small liberal arts school, Wheaton College in Massachusetts, devised a strategic plan to improve faculty salaries by linking improvements in salaries to improvements in college resources. The benefits of the salary plan went beyond improving salaries. First, the plan dictated that faculty salaries be determined at…

  17. Advanced strategic missile development

    NASA Astrophysics Data System (ADS)

    Strickler, R. L.

    1981-05-01

    The M-X program is taking two paths: (1) the current development and projected deployment of a survivable land based ICBM (the M-X) in a multiple protective structure system, and (2) a building block development of readiness posture and strategic futures technology that could be used for a wide range of projected needs in the event of major changes in the threat or the political climate. The blend of aerospace and civil engineering technologies which has resulted in the systems concept necessary to assure the continued survivability of the land based strategic missile force is summarized. Recent advanced technology development activities, which have been focused on systems upgrade options to the current ICBM force, basing options which may be required for special force elements, small missile options for airborne applications, penetration technology to counter SAM and ABM threats, and systems concepts for unique targeting requirements are reviewed.

  18. Pain Raises Risk of Opioid Addiction

    MedlinePlus

    ... fullstory_160033.html Pain Raises Risk of Opioid Addiction Men and younger people had higher odds of ... had a 41 percent higher risk of opioid addiction than those with no pain. That increased risk ...

  19. 7 CFR 60.123 - Raised.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...) Farm-raised fish and shellfish as it relates to the production steps defined in § 60.122: The period of... MARKETING ACT OF 1946 AND THE EGG PRODUCTS INSPECTION ACT (CONTINUED) COUNTRY OF ORIGIN LABELING FOR...

  20. 7 CFR 60.123 - Raised.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...) Farm-raised fish and shellfish as it relates to the production steps defined in § 60.122: The period of... MARKETING ACT OF 1946 AND THE EGG PRODUCTS INSPECTION ACT (CONTINUED) COUNTRY OF ORIGIN LABELING FOR...

  1. Pesticides Linked to Raised Risk of ALS

    MedlinePlus

    ... nlm.nih.gov/medlineplus/news/fullstory_158737.html Pesticides Linked to Raised Risk of ALS One toxin ... MONDAY, May 9, 2016 (HealthDay News) -- Exposure to pesticides and other chemicals may increase the risk for ...

  2. Alcohol, Obesity Could Raise Esophageal Cancer Risk

    MedlinePlus

    ... https://medlineplus.gov/news/fullstory_160133.html Alcohol, Obesity Could Raise Esophageal Cancer Risk A third of ... at the American Institute for Cancer Research (AICR). "Obesity is now linked to 11 types of cancer ...

  3. Common Surgeries Raise Risk for Opioid Dependence

    MedlinePlus

    ... fullstory_159815.html Common Surgeries Raise Risk for Opioid Dependence: Study Doctors should explore alternatives for pain ... have an elevated risk of growing dependent on opioid painkillers, a new study finds. These prescription painkillers ...

  4. Pesticides Linked to Raised Risk of ALS

    MedlinePlus

    ... page: https://medlineplus.gov/news/fullstory_158737.html Pesticides Linked to Raised Risk of ALS One toxin ... MONDAY, May 9, 2016 (HealthDay News) -- Exposure to pesticides and other chemicals may increase the risk for ...

  5. The Strategic Petroleum Reserve

    SciTech Connect

    Not Available

    1991-01-01

    The Strategic Petroleum Reserve program was set into motion by the 1975 Energy Policy and Conservation Act (EPCA). By 1990, 590 million barrels of oil had been placed in storage. Salt domes along the Gulf Coast offered ideal storage. Both sweet'' and sour'' crude oil have been acquired using various purchase options. Drawdown, sale, and distribution of the oil would proceed according to guidelines set by EPCA in the event of a severe energy supply disruption. (SM)

  6. Microgravity strategic plan, 1988

    NASA Technical Reports Server (NTRS)

    1988-01-01

    The NASA agency-wide microgravity strategic plan is presented, and its research, applications, and commercialization for the 1990's is addressed. The plan presents an analysis of the current situation, identifies critical factors, and defines goals, objectives, and strategies, which are intended to: (1) provide a context for decision making; (2) assure realism in long-range planning and direction for hardware development; and (3) establish a framework for developing a national microgravity research plan.

  7. Strategic plan, 1985

    NASA Technical Reports Server (NTRS)

    1985-01-01

    The Lewis Strategic Plan was updated for 1985 and beyond. Major programs for the space station, the advanced turboprop, the Advanced Communications Technology Satellite (ACTS), and the Altitude Wind Tunnel were begun or greatly expanded during 1984. In parallel, The Lewis aeropropulsion research and technology program was extensively evaluated and reviewed; a reduced and reoriented program emerged. The thrusts and implementation plans for these programs are described as they pertain to the individual directorates. Other key accomplishments and plans are summarized.

  8. Generation of ER{alpha}-floxed and knockout mice using the Cre/LoxP system

    SciTech Connect

    Antonson, P.; Omoto, Y.; Humire, P.; Gustafsson, J.-A.

    2012-08-10

    Highlights: Black-Right-Pointing-Pointer ER{alpha} floxed and knockout mice were generated. Black-Right-Pointing-Pointer Disruption of the ER{alpha} gene results in sterility in both male and female mice. Black-Right-Pointing-Pointer ER{alpha}{sup -/-} mice have ovaries with hemorrhagic follicles and hypoplastic uterus. Black-Right-Pointing-Pointer Female ER{alpha}{sup -/-} mice develop obesity. -- Abstract: Estrogen receptor alpha (ER{alpha}) is a nuclear receptor that regulates a range of physiological processes in response to estrogens. In order to study its biological role, we generated a floxed ER{alpha} mouse line that can be used to knock out ER{alpha} in selected tissues by using the Cre/LoxP system. In this study, we established a new ER{alpha} knockout mouse line by crossing the floxed ER{alpha} mice with Cre deleter mice. Here we show that genetic disruption of the ER{alpha} gene in all tissues results in sterility in both male and female mice. Histological examination of uterus and ovaries revealed a dramatically atrophic uterus and hemorrhagic cysts in the ovary. These results suggest that infertility in female mice is the result of functional defects of the reproductive tract. Moreover, female knockout mice are hyperglycemic, develop obesity and at the age of 4 months the body weight of these mice was more than 20% higher compared to wild type littermates and this difference increased over time. Our results demonstrate that ER{alpha} is necessary for reproductive tract development and has important functions as a regulator of metabolism in females.

  9. Viral transduction of primary Schwann cells using a Cre-lox system to regulate GDNF expression.

    PubMed

    Wu-Fienberg, Yuewei; Moore, Amy M; Marquardt, Laura M; Newton, Piyaraj; Johnson, Philip J; Mackinnon, Susan E; Sakiyama-Elbert, Shelly E; Wood, Matthew D

    2014-09-01

    Glial cell-line-derived neurotrophic factor (GDNF) is a potent neurotrophic factor known to enhance motor nerve regeneration following its delivery. However, recent studies have determined that extended GDNF delivery to regenerating axons can entrap motor axons at the site of GDNF delivery. This entrapment leads to reduced motor axons available to reinnervate muscle. To address this issue, we designed a cell-based GDNF expression system that can temporally regulate protein expression using an inducible gene excision mechanism to prevent entrapment at the site of expression. To design this system for regulation of GDNF expression, we transduced two lentiviral vectors, one containing a constitutively active GDNF transgene flanked by two loxP sites, and the other containing a tetracycline-inducible cre transgene along with its constitutively active transactivator, into Schwann cells (SCs). These SCs over-express GDNF, but expression can be suppressed through the administration of tetracycline family antibiotics, such as doxycycline. The engineered SCs produced significantly more GDNF as compared to untransduced controls, as measured by enzyme-linked immunosorbent assay (ELISA). Following doxycycline treatment, these SCs produced significantly lower levels of GDNF and induced less neurite extension as compared to untreated SCs. Engineered SCs treated with doxycycline showed a marked increase in Cre recombinase expression, as visualized by immunohistochemistry (IHC), providing evidence of a mechanism for the observed changes in GDNF expression levels and biological activity. This cell-based GDNF expression system could have potential for future in vivo studies to provide a temporally controlled GDNF source to promote axon growth. PMID:24728940

  10. Viral transduction of primary Schwann cells using a Cre-lox system to regulate GDNF expression

    PubMed Central

    Wu-Fienberg, Yuewei; Moore, Amy M.; Marquardt, Laura M.; Newton, Piyaraj; Johnson, Philip J.; Mackinnon, Susan E.; Sakiyama-Elbert, Shelly E.; Wood, Matthew D.

    2014-01-01

    Glial cell-line-derived neurotrophic factor (GDNF) is a potent neurotrophic factor known to enhance motor nerve regeneration following its delivery. However, recent studies have determined that extended GDNF delivery to regenerating axons can entrap motor axons at the site of GDNF delivery. This entrapment leads to reduced motor axons available to reinnervate muscle. To address this issue, we designed a cell-based GDNF expression system that can temporally regulate protein expression using an inducible gene excision mechanism to prevent entrapment at the site of expression. To design this system for regulation of GDNF expression, we transduced two lentiviral vectors, one containing a constitutively active GDNF transgene flanked by two loxP sites, and the other containing a tetracycline-inducible cre transgene along with its constitutively active transactivator, into Schwann cells (SCs). These SCs over-express GDNF, but expression can be suppressed through the administration of tetracycline family antibiotics, such as doxycycline. The engineered SCs produced significantly more GDNF as compared to untransduced controls, as measured by enzyme-linked immunosorbent assay (ELISA). Following doxycycline treatment, these SCs produced significantly lower levels of GDNF and induced less neurite extension as compared to untreated SCs. Engineered SCs treated with doxycycline showed a marked increase in Cre recombinase expression, as visualized by immunohistochemistry (IHC), providing evidence of a mechanism for the observed changes in GDNF expression levels and biological activity. This cell-based GDNF expression system could have potential for future in vivo studies to provide a temporally-controlled GDNF source to promote axon growth. PMID:24728940

  11. Vegfr3-CreER (T2) mouse, a new genetic tool for targeting the lymphatic system.

    PubMed

    Martinez-Corral, Ines; Stanczuk, Lukas; Frye, Maike; Ulvmar, Maria Helena; Diéguez-Hurtado, Rodrigo; Olmeda, David; Makinen, Taija; Ortega, Sagrario

    2016-07-01

    The lymphatic system is essential in many physiological and pathological processes. Still, much remains to be known about the molecular mechanisms that control its development and function and how to modulate them therapeutically. The study of these mechanisms will benefit from better controlled genetic mouse models targeting specifically lymphatic endothelial cells. Among the genes expressed predominantly in lymphatic endothelium, Vegfr3 was the first one identified and is still considered to be one of the best lymphatic markers and a key regulator of the lymphatic system. Here, we report the generation of a Vegfr3-CreER (T2) knockin mouse by gene targeting in embryonic stem cells. This mouse expresses the tamoxifen-inducible CreER(T2) recombinase under the endogenous transcriptional control of the Vegfr3 gene without altering its physiological expression or regulation. The Vegfr3-CreER (T2) allele drives efficient recombination of floxed sequences upon tamoxifen administration specifically in Vegfr3-expressing cells, both in vitro, in primary lymphatic endothelial cells, and in vivo, at different stages of mouse embryonic development and postnatal life. Thus, our Vegfr3-CreER (T2) mouse constitutes a new powerful genetic tool for lineage tracing analysis and for conditional gene manipulation in the lymphatic endothelium that will contribute to improve our current understanding of this system. PMID:26993803

  12. Diminished CRE-Induced Plasticity is Linked to Memory Deficits in Familial Alzheimer’s Disease Mice

    PubMed Central

    Bartolotti, Nancy; Segura, Laura; Lazarov, Orly

    2015-01-01

    The mechanism underlying impaired learning and memory in Alzheimer’s disease is not fully elucidated. The phosphorylation of cyclic-AMP response element binding protein (pCREB) in the hippocampus is thought to be a critical initiating step in the formation of long-term memories. Here, we tested CRE-driven gene expression following learning in mice harboring the familial Alzheimer’s disease-linked APPswe/PS1ΔE9 mutations using CRE-β galactosidase reporter. We show that young adult APPswe/PS1ΔE9 mice exhibit impaired recognition memory and reduced levels of pCREB, and its cofactors CREB binding protein (CBP) and p-300 following a learning task, compared to their wild type littermate counterparts. Impairments in learning-induced activation of CREB in these mice are manifested by reduced CRE-driven gene transcription. Importantly, expression of the CRE-driven immediate early gene, Egr-1 (Zif268) is decreased in the CA1 region of the hippocampus. These studies implicate defective CREB-dependent plasticity in the mechanism underlying learning and memory deficits in Alzheimer’s disease. PMID:26682682

  13. Cre/lox system to develop selectable marker free transgenic tobacco plants conferring resistance against sap sucking homopteran insect.

    PubMed

    Chakraborti, Dipankar; Sarkar, Anindya; Mondal, Hossain A; Schuermann, David; Hohn, Barbara; Sarmah, Bidyut K; Das, Sampa

    2008-10-01

    A binary expression vector was constructed containing the insecticidal gene Allium sativum leaf agglutinin (ASAL), and a selectable nptII marker gene cassette, flanked by lox sites. Similarly, another binary vector was developed with the chimeric cre gene construct. Transformed tobacco plants were generated with these two independent vectors. Each of the T(0) lox plants was crossed with T(0) Cre plants. PCR analyses followed by the sequencing of the target T-DNA part of the hybrid T(1) plants demonstrated the excision of the nptII gene in highly precised manner in certain percentage of the T(1) hybrid lines. The frequency of such marker gene excision was calculated to be 19.2% in the hybrids. Marker free plants were able to express ASAL efficiently and reduce the survivability of Myzus persiceae, the deadly pest of tobacco significantly, compared to the control tobacco plants. Results of PCR and Southern blot analyses of some of the T(2) plants detected the absence of cre as well as nptII genes. Thus, the crossing strategy involving Cre/lox system for the excision of marker genes appears to be very effective and easy to execute. Documentation of such marker excision phenomenon in the transgenic plants expressing the important insecticidal protein for the first time has a great significance from agricultural and biotechnological points of view. PMID:18663453

  14. A Cre-conditional MYCN-driven neuroblastoma mouse model as an improved tool for preclinical studies.

    PubMed

    Althoff, K; Beckers, A; Bell, E; Nortmeyer, M; Thor, T; Sprüssel, A; Lindner, S; De Preter, K; Florin, A; Heukamp, L C; Klein-Hitpass, L; Astrahantseff, K; Kumps, C; Speleman, F; Eggert, A; Westermann, F; Schramm, A; Schulte, J H

    2015-06-01

    Neuroblastoma, a childhood cancer that originates from neural crest-derived cells, is the most common deadly solid tumor of infancy. Amplification of the MYCN oncogene, which occurs in approximately 20-25% of human neuroblastomas, is the most prominent genetic marker of high-stage disease. The availability of valid preclinical in vivo models is a prerequisite to develop novel targeted therapies. We here report on the generation of transgenic mice with Cre-conditional induction of MYCN in dopamine β-hydroxylase-expressing cells, termed LSL-MYCN;Dbh-iCre. These mice develop neuroblastic tumors with an incidence of >75%, regardless of strain background. Molecular profiling of tumors revealed upregulation of the MYCN-dependent miR-17-92 cluster as well as expression of neuroblastoma marker genes, including tyrosine hydroxylase and the neural cell adhesion molecule 1. Gene set enrichment analyses demonstrated significant correlation with MYC-associated expression patterns. Array comparative genome hybridization showed that chromosomal aberrations in LSL-MYCN;Dbh-iCre tumors were syntenic to those observed in human neuroblastomas. Treatment of a cell line established from a tumor derived from a LSL-MYCN;Dbh-iCre mouse with JQ1 or MLN8237 reduced cell viability and demonstrated oncogene addiction to MYCN. Here we report establishment of the first Cre-conditional human MYCN-driven mouse model for neuroblastoma that closely recapitulates the human disease with respect to tumor localization, histology, marker expression and genomic make up. This mouse model is a valuable tool for further functional studies and to assess the effect of targeted therapies. PMID:25174395

  15. Strategic Petroleum Reserve quarterly report

    SciTech Connect

    Not Available

    1990-08-15

    The Strategic Petroleum Reserve Quarterly Report is submitted in accordance with section 165(b) of the Energy Policy and Conservation Act, as amended, which requires that the Secretary of Energy submit quarterly reports to Congress on Activities undertaken with respect to the Strategic Petroleum Reserve. This August 15, 1990, Strategic Petroleum Reserve Quarterly Report describes activities related to the site development, oil acquisition, budget and cost of the Reserve during the period April 1, 1990, through June 30, 1990. 3 tabs.

  16. National strategic petroleum reserve.

    PubMed

    Davis, R M

    1981-08-01

    The Strategic Petroleum Reserve is intended to reduce the vulnerability of the United States to interruptions in the oil supply from foreign sources. Storage for 248 million barrels of crude oil in salt caverns and mines, with equipment for pumping and distribution, was constructed and operationally tested in a 4-year period. Its present inventory is the largest known crude oil reserve in the world. Facilities for expanding the reserve's capacity by another 290 million barrels are being developed by solution-mining in salt domes. PMID:17847458

  17. NASA strategic plan

    NASA Technical Reports Server (NTRS)

    1995-01-01

    NASA's Plan summarizes the Agency's vision, mission, and values. Specific goals are listed for each externally focused Enterprise: Mission to Planet Earth, Aeronautics, Human Exploration and Development of Space, Space Science, and Space Technology. These Enterprises satisfy the needs of customers external to NASA. The Strategic Functions (Space Communications, Human Resources, and Physical Resources) are necessary in order to meet the goals of the Enterprises. The goals of these Functions are also presented. All goals must be met while adhering to the discussed values and operating principles of NASA. A final section outlines the implementing strategy.

  18. Guam Strategic Energy Plan

    SciTech Connect

    Conrad, M. D.

    2013-07-01

    Describes various energy strategies available to Guam to meet the territory's goal of diversifying fuel sources and reducing fossil energy consumption 20% by 2020.The information presented in this strategic energy plan will be used by the Guam Energy Task Force to develop an energy action plan. Available energy strategies include policy changes, education and outreach, reducing energy consumption at federal facilities, and expanding the use of a range of energy technologies, including buildings energy efficiency and conservation, renewable electricity production, and alternative transportation. The strategies are categorized based on the time required to implement them.

  19. Strategic implementation plan

    NASA Technical Reports Server (NTRS)

    1989-01-01

    The Life Science Division of the NASA Office of Space Science and Applications (OSSA) describes its plans for assuring the health, safety, and productivity of astronauts in space, and its plans for acquiring further fundamental scientific knowledge concerning space life sciences. This strategic implementation plan details OSSA's goals, objectives, and planned initiatives. The following areas of interest are identified: operational medicine; biomedical research; space biology; exobiology; biospheric research; controlled ecological life support; flight programs and advance technology development; the life sciences educational program; and earth benefits from space life sciences.

  20. Reversed DNA strand cleavage specificity in initiation of Cre-LoxP recombination induced by the His289Ala active-site substitution.

    PubMed

    Gelato, Kathy A; Martin, Shelley S; Baldwin, Enoch P

    2005-11-25

    During the first steps of site-specific recombination, Cre protein cleaves and religates a specific homologous pair of LoxP strands to form a Holliday junction (HJ) intermediate. The HJ is resolved into recombination products through exchange of the second homologous strand pair. CreH289A, containing a His to Ala substitution in the conserved R-H-R catalytic motif, has a 150-fold reduced recombination rate and accumulates HJs. However, to produce these HJs, CreH289A exchanges the opposite set of strands compared to wild-type Cre (CreWT). To investigate how CreH289A and CreWT impose strand exchange order, we characterized their reactivities and strand cleavage preferences toward LoxP duplex and HJ substrates containing 8bp spacer substitutions. Remarkably, CreH289A had different and often opposite strand exchange preferences compared to CreWT with nearly all substrates. CreH289N was much less perturbed, implying that overall recombination rate and strand exchange depend more on His289 hydrogen bonding capability than on its acid/base properties. LoxP substitutions immediately 5' (S1 nucleotide) or 3' (S1' nucleotide) of the scissile phosphate had large effects on substrate utilization and strand exchange order. S1' substitutions, designed to alter base-unstacking events concomitant with Cre-induced LoxP bending, caused HJ accumulation and dramatically inverted the cleavage preferences. That pre-formed HJs were resolved via either strand in vitro suggests that inhibition of the "conformational switch" isomerization required to trigger the second strand exchange accounts for the observed HJ accumulation. Rather than reflecting CreWT behavior, CreH289A accumulates HJs of opposite polarity through a combination of its unique cleavage specificity and an HJ isomerization defect. The overall implication is that cleavage specificity is mediated by sequence-dependent DNA deformations that influence the scissile phosphate positioning and reactivity. A role of His289 may be to

  1. Strategic Responses to Financial Challenges. Strategic Decisions. Board Basics.

    ERIC Educational Resources Information Center

    Hauptman, Arthur M.

    1998-01-01

    This booklet for trustees of institutions of higher education offers guidelines for strategic financial decisions required in light of the financial challenges higher education faces in the 1990s and beyond. "Strategic response" is defined as a decision that requires changing a major policy, program, or practice and involves some risk. Five…

  2. Strategic Decisions in Task-Oriented Reading.

    PubMed

    Salmerón, Ladislao; Vidal-Abarca, Eduardo; Martínez, Tomás; Mañá, Amelia; Gil, Laura; Naumann, Johannes

    2015-01-01

    Answering questions from texts are assessment and instructional activities that are frequently used in schools. Nevertheless, little is known about the strategic processes that students take while performing these tasks. We explored the amount and frequency that students initially read of a text before they answered questions pertaining to the material. In a procedure similar to the one used in the PISA (Program for International Students Assessment), one-hundred-seventy students between 7th and 9th grade read and answered several questions designed to assess task-oriented reading in three specific texts. We recorded on-line indexes that evaluated student behavior (e.g., the amount of text that students read before answering questions raised within a given text), performance, and comprehension skill. The results revealed that students skilled in comprehension initially read a high proportion of the texts, which in turn improved their overall performance in two of the three texts read (text 1: CI95%: 0.01 to 0.09; text 2: CI95%: -0.01 to 0.05; text 3: CI95%: 0.04 to 0.20). Therefore, we conclude that this strategic behavior should be considered during the assessment and instruction of reading literacy. PMID:26707969

  3. CRE dating on the scarps of large landslides affecting the Belledonne massif ( French Alps)

    NASA Astrophysics Data System (ADS)

    Lebrouc, V.; Baillet, L.; Schwartz, S.; Jongmans, D.; Gamond, J. F.; Bourles, D.; Le Roux, O.; Carcaillet, J.; Braucher, R.

    2012-04-01

    The southwestern edge of the Belledonne Massif (French Alps) consists of micaschists unconformably covered with Mesozoic sediments and Quaternary deposits. The morphology corresponds to a glacial plateau (Mont Sec plateau) bordered by steep slopes (around 40°), where moraines and peat bog subsist. The massif is incised by the East-West trending Romanche valley that was shaped by several cycles of quaternary glaciations and deglaciations. Slopes are affected by several active or past large scale rock mass instabilities. Cosmic Ray Exposure (CRE) dating was applied on the head scarps of three large landslides, one of which being the active Séchilienne landslide whose headscarp was already dated by Leroux et al. [2009]. Dating results suggest a concomitant initiation of these instabilities at about 7 ± 2 10Be ka, thousands years after the total downwastage of the valley. A different kinematic behaviour was however observed on two contiguous landslides for which continuous exposure profiles were obtained. On the Séchilienne landslide, 23 samples were collected from internal and lateral scarps, as well as on polished bedrock surfaces, with the aim of dating the internal kinematics of the landslide. Preliminary dating results obtained on polished surfaces and near the top of the scarps show unexpected low 10Be concentrations, suggesting the existence of thin moraine or peat bog deposits masking the bedrock, which have been subsequently eroded. The minimum thickness of these deposits was estimated assuming a constant denudation rate over time. Exposure date profiles show that the studied lateral and internal scarps were initiated at the same period as the Sechilienne headscarp. An increase in the exposure rate was also observed between 2 and 1 ka, in agreement with that evidenced along the headscarp. Forty other samples have been collected in the landslide to corroborate these results. Reference Le Roux, O., S. Schwartz , J.-F. Gamond, D. Jongmans, D. Bourles, R

  4. The CRE1 Cytokinin Pathway Is Differentially Recruited Depending on Medicago truncatula Root Environments and Negatively Regulates Resistance to a Pathogen

    PubMed Central

    Laffont, Carole; Rey, Thomas; André, Olivier; Novero, Mara; Kazmierczak, Théophile; Debellé, Frédéric; Bonfante, Paola; Jacquet, Christophe; Frugier, Florian

    2015-01-01

    Cytokinins are phytohormones that regulate many developmental and environmental responses. The Medicago truncatula cytokinin receptor MtCRE1 (Cytokinin Response 1) is required for the nitrogen-fixing symbiosis with rhizobia. As several cytokinin signaling genes are modulated in roots depending on different biotic and abiotic conditions, we assessed potential involvement of this pathway in various root environmental responses. Phenotyping of cre1 mutant roots infected by the Gigaspora margarita arbuscular mycorrhizal (AM) symbiotic fungus, the Aphanomyces euteiches root oomycete, or subjected to an abiotic stress (salt), were carried out. Detailed histological analysis and quantification of cre1 mycorrhized roots did not reveal any detrimental phenotype, suggesting that MtCRE1 does not belong to the ancestral common symbiotic pathway shared by rhizobial and AM symbioses. cre1 mutants formed an increased number of emerged lateral roots compared to wild-type plants, a phenotype which was also observed under non-stressed conditions. In response to A. euteiches, cre1 mutants showed reduced disease symptoms and an increased plant survival rate, correlated to an enhanced formation of lateral roots, a feature previously linked to Aphanomyces resistance. Overall, we showed that the cytokinin CRE1 pathway is not only required for symbiotic nodule organogenesis but also affects both root development and resistance to abiotic and biotic environmental stresses. PMID:25562779

  5. Second-generation Notch1 activity-trap mouse line (N1IP::CreHI) provides a more comprehensive map of cells experiencing Notch1 activity

    PubMed Central

    Liu, Zhenyi; Brunskill, Eric; Boyle, Scott; Chen, Shuang; Turkoz, Mustafa; Guo, Yuxuan; Grant, Rachel; Kopan, Raphael

    2015-01-01

    We have previously described the creation and analysis of a Notch1 activity-trap mouse line, Notch1 intramembrane proteolysis-Cre6MT or N1IP::CreLO, that marked cells experiencing relatively high levels of Notch1 activation. Here, we report and characterize a second line with improved sensitivity (N1IP::CreHI) to mark cells experiencing lower levels of Notch1 activation. This improvement was achieved by increasing transcript stability and by restoring the native carboxy terminus of Cre, resulting in a five- to tenfold increase in Cre activity. The magnitude of this effect probably impacts Cre activity in strains with carboxy-terminal Ert2 fusion. These two trap lines and the related line N1IP::CreERT2 form a complementary mapping tool kit to identify changes in Notch1 activation patterns in vivo as the consequence of genetic or pharmaceutical intervention, and illustrate the variation in Notch1 signal strength from one tissue to the next and across developmental time. PMID:25725069

  6. Second-generation Notch1 activity-trap mouse line (N1IP::CreHI) provides a more comprehensive map of cells experiencing Notch1 activity.

    PubMed

    Liu, Zhenyi; Brunskill, Eric; Boyle, Scott; Chen, Shuang; Turkoz, Mustafa; Guo, Yuxuan; Grant, Rachel; Kopan, Raphael

    2015-03-15

    We have previously described the creation and analysis of a Notch1 activity-trap mouse line, Notch1 intramembrane proteolysis-Cre6MT or N1IP::Cre(LO), that marked cells experiencing relatively high levels of Notch1 activation. Here, we report and characterize a second line with improved sensitivity (N1IP::Cre(HI)) to mark cells experiencing lower levels of Notch1 activation. This improvement was achieved by increasing transcript stability and by restoring the native carboxy terminus of Cre, resulting in a five- to tenfold increase in Cre activity. The magnitude of this effect probably impacts Cre activity in strains with carboxy-terminal Ert2 fusion. These two trap lines and the related line N1IP::Cre(ERT2) form a complementary mapping tool kit to identify changes in Notch1 activation patterns in vivo as the consequence of genetic or pharmaceutical intervention, and illustrate the variation in Notch1 signal strength from one tissue to the next and across developmental time. PMID:25725069

  7. Raising Butterflies from Your Own Garden.

    ERIC Educational Resources Information Center

    Howley-Pfeifer, Patricia

    2002-01-01

    Describes how raising monarch, black swallowtail, and mourning cloak butterflies in a kindergarten class garden can provide opportunities for observation experiences. Includes detailed steps for instruction and describes stages of growth. Excerpts children's journal dictations to illustrate ways to support the discovery process. Describes related…

  8. People on the Farm: Raising Beef Cattle.

    ERIC Educational Resources Information Center

    Crain, Robert L.

    This booklet provides information on raising beef cattle through profiles of two families, the Ritschards of Colorado and the Schuttes of Missouri. Through descriptions of daily life for these families, the booklet discusses the way of life on modern beef cattle farms and the problems and decisions faced by farmers. The booklet explains how…

  9. Infant Botulism and Raised Intraocular Pressure.

    PubMed

    Eberly, Matthew D; Uber, Ian; Kieling, Christopher R; Birdsong, Richard H

    2009-11-01

    Infant botulism is an exceedingly rare disease. Because confirmatory laboratory testing is not available for several days after time of presentation, infant botulism remains a clinical diagnosis. The authors demonstrate how raised intraocular pressure may provide an additional clinical clue to making the diagnosis. PMID:19873952

  10. YOU CAN RAISE YOUR HANDICAPPED CHILD.

    ERIC Educational Resources Information Center

    AYRAULT, EVELYN WEST

    DESIGNED AS A GUIDE FOR PARENTS OF HANDICAPPED CHILDREN, THIS BOOK PROVIDES INFORMATION ON TYPES OF HANDICAPS AND THE AIMS AND TECHNIQUES OF TREATMENT, THERAPY, AND EDUCATION. SPECIFIC RECOMMENDATIONS TO HELP PARENTS RAISE A HANDICAPPED CHILD ARE DISCUSSED. BEHAVIOR PROBLEMS, PARENT ATTITUDES, DAILY PROBLEMS, AND RECREATION ARE EXAMINED IN DETAIL…

  11. Consciousness-Raising, Error Correction and Proofreading

    ERIC Educational Resources Information Center

    O'Brien, Josephine

    2015-01-01

    The paper discusses the impact of developing a consciousness-raising approach in error correction at the sentence level to improve students' proofreading ability. Learners of English in a foreign language environment often rely on translation as a composing tool and while this may act as a scaffold and provide some support, it frequently leads to…

  12. Raising Happy Kids on a Reasonable Budget.

    ERIC Educational Resources Information Center

    Gallagher, Patricia C.

    This guide shows parents how to cut expenses and save on the cost of raising a family. Chapter 1 offers many ways to stretch food dollars and discusses buying in bulk, eating out, using coupons, buying wholesale, and gardening. Cost-cutting clothing ideas covered in chapter 2 include buying at secondhand stores and consignment shops, outlet…

  13. Winning Techniques for Athletic Fund Raising.

    ERIC Educational Resources Information Center

    Alberger, Patti, Ed.

    Techniques are presented from successful athletic fund-raisers on how to structure a fund-raising program through an independent booster club, as part of the athletic department or as part of the total institutional development program. The handbook offers advice on building a program from scratch; organizing successful campaigns, involving…

  14. Raising Environmental Awareness among Miners in Iran

    ERIC Educational Resources Information Center

    Mozaffari, Ezatollah

    2013-01-01

    Generation of waste is inevitable but controllable in minerals industry. The aim of this research is to find ways for raising environmental awareness among miners. Miners' attitude towards environmental mining has been investigated. A survey has been done collecting mine managers' point of view coupled with current trend on mine waste management…

  15. Prospects for College and University Fund Raising.

    ERIC Educational Resources Information Center

    Hoeflich, M. H.

    1987-01-01

    Changes in fund raising because of the implementation of the 1986 Tax Reform Act are discussed. The impact on individual and corporate donors is described. Universities and colleges must increase their efforts on the state level to assure themselves a fair share of any windfall revenues and bond-issuance authority. (MLW)

  16. Fund Raising in ARL Libraries. Kit 94.

    ERIC Educational Resources Information Center

    Association of Research Libraries, Washington, DC. Office of Management Studies.

    In January 1983 the Association of Research Libraries (ARL) surveyed its members regarding their fund raising programs. A response rate of 85% (95 out of 113 libraries surveyed) was obtained. It was found that 85% of ARL member libraries had been involved in university-wide fund campaigns; that over 75% of the respondents had also conducted…

  17. Solo Parenting: Raising Strong & Happy Families.

    ERIC Educational Resources Information Center

    Chambers, Diane

    One quarter of all American children will live in a single-parent family for at least some portion of their childhood. This guide, organized as a step-by-step process, encourages single parents to reach for excellence in their parenting and to build confidence in their ability to raise healthy, responsible children. The chapters are intended to…

  18. Raising Literacy Standards in the Primary Division.

    ERIC Educational Resources Information Center

    Waiser, Marlene

    A research project undertaken in York, Ontario, Canada aimed to raise literacy standards in the primary division and to ensure that all children in Grade 1 read to the Grade 1 standard (as determined by Reading Recovery levels, the school board, and the Ontario Ministry of Education). Focus was placed on acquiring high quality leveled literature…

  19. Liberation Therapeutics: Consciousness Raising as a Problem.

    ERIC Educational Resources Information Center

    Lasch-Quinn, Elisabeth

    2002-01-01

    Questions the content and form of consciousness raising as a mode of purveying knowledge or bringing about change by considering its emergence in the civil rights movement. Examines such books as "Black Rage" (William Grierand Price Cobbs), "Triumph of the Therapeutic" (Philip Reiff), "Where Do We Go From Here: Chaos or Community?" (Martin Luther…

  20. The Fundamentals of Community College Fund Raising.

    ERIC Educational Resources Information Center

    Brumbach, Mary A.; Bumphus, Walter G.

    1993-01-01

    Presents a guide for community college fund raising. Discusses organizational principles and structural models; operational concerns such as stewardship of funds, information dissemination, and donor research; and the importance of presidential support, institutional reputation and environment, and experienced grants managers. Includes practical…

  1. National Organizations Raising Public Consciousness. Part II.

    ERIC Educational Resources Information Center

    Exceptional Parent, 1980

    1980-01-01

    Statements of seven national organizations (Muscular Dystrophy Association, Spina Bifida Association of America, National Easter Seal Society, International Association of Parents of the Deaf, American Foundation for the Blind, Down's Syndrome Congress, and the National Society for Autistic Children) about raising community awareness of issues…

  2. Guidelines Manual for Fund Raising and Donations.

    ERIC Educational Resources Information Center

    Tidewater Community Coll. Educational Foundation, Inc., Portsmouth, VA.

    Guidelines and procedures for fund raising and the acceptance of gifts at Tidewater Community College (TCC) are presented in this manual. First, general information is provided on the acceptance of gifts of cash, securities, real estate, insurance, personal property, equipment, and gifts requiring the expenditure of funds. Next, general procedures…

  3. Raising Money Through an Institutionally Related Foundation.

    ERIC Educational Resources Information Center

    Reilley, Timothy A., Ed.

    The creation of foundations for fund raising at public colleges and new ideas and techniques for established foundations are discussed in 13 chapters. The relationship of the foundation and the institution is described from the viewpoint of the institution and also that of the foundation. Article titles and authors include: "How the Foundation…

  4. Children Seem to Know Raising: Raising and Intervention in Child Language

    ERIC Educational Resources Information Center

    Choe, Jinsun

    2012-01-01

    English-speaking children exhibit difficulty in their comprehension of raising patterns, such as (1), in which the NP the boy is semantically linked to the VP in the embedded clause, but is syntactically realized as the subject of the matrix clause. (1) Raising pattern: [s "The boy" seems to the girl [s _ to be happy

  5. Strategic Planning Is an Oxymoron

    ERIC Educational Resources Information Center

    Bassett, Patrick F.

    2012-01-01

    The thinking on "strategic thinking" has evolved significantly over the years. In the previous century, the independent school strategy was to focus on long-range planning, blithely projecting 10 years into the future. For decades this worked well enough, but in the late 20th century, independent schools shifted to "strategic planning," with its…

  6. Strategic Partnerships in International Development

    ERIC Educational Resources Information Center

    Treat, Tod; Hartenstine, Mary Beth

    2013-01-01

    This chapter provides a framework and recommendations for development of strategic partnerships in a variety of cultural contexts. Additionally, this study elucidates barriers and possibilities in interagency collaborations. Without careful consideration regarding strategic partnerships' approaches, functions, and goals, the ability to…

  7. The Search for Strategic Silence.

    ERIC Educational Resources Information Center

    Lentz, Richard

    Media content analysts seldom observe the principle that editorial omissions are as telling as what is published or broadcast; hence, the purpose of this paper is to explore, and thus stimulate debate about, editorial omissions or "strategic silence." It is observed that as a concept, strategic silence embraces both tact and strategy--the former…

  8. NASA Space Sciences Strategic Planning

    NASA Technical Reports Server (NTRS)

    Crane, Philippe

    2004-01-01

    The purpose of strategic planning roadmap is to:Fulfill the strategic planning requirements; Provide a guide to the science community in presenting research requests to NASA; Inform and inspire; Focus investments in technology and research for future missions; and Provide the scientific and technical justification for augmentation requests.

  9. Strategic Human Resource Development. Symposium.

    ERIC Educational Resources Information Center

    2002

    This document contains three papers on strategic human resource (HR) development. "Strategic HR Orientation and Firm Performance in India" (Kuldeep Singh) reports findings from a study of Indian business executives that suggests there is a positive link between HR policies and practices and workforce motivation and loyalty and sustainable…

  10. Strategic planning for the board.

    PubMed

    Davies, A

    1991-04-01

    Although the planning operation is regarded by some observers as unrealistic in conditions of rapid change and increasing competition, the discipline of strategic thinking and the need for strategic leadership continue to be of vital importance. The author examines the purpose of the Board of Directors and its role in the management of strategy. PMID:10111271

  11. Strategic Evaluation in University Management.

    ERIC Educational Resources Information Center

    Tavernier, Karel

    1991-01-01

    Improved evaluation of higher education teaching and research is urged, citing need for quantitative and qualitative indicators of an institution's strategic capacity to adapt to external demands and for closer links between evaluation and institutional goals. Industry's strategic management methods and difficulties of evaluation in different…

  12. Strategic Marketing for Educational Systems.

    ERIC Educational Resources Information Center

    Hanson, E. Mark; Henry, Walter

    1992-01-01

    Private-sector strategic marketing processes can significantly benefit schools desiring to develop public confidence and support and establish guidelines for future development. This article defines a strategic marketing model for school systems and articulates the sequence of related research and operational steps comprising it. Although schools…

  13. Strategic Planning for Higher Education.

    ERIC Educational Resources Information Center

    Kotler, Philip; Murphy, Patrick E.

    1981-01-01

    The framework necessary for achieving a strategic planning posture in higher education is outlined. The most important benefit of strategic planning for higher education decision makers is that it forces them to undertake a more market-oriented and systematic approach to long- range planning. (Author/MLW)

  14. Developing the Strategically Focused School

    ERIC Educational Resources Information Center

    Davies, Brent

    2004-01-01

    This article builds a conceptual framework to examine how school leaders can move away from a planning framework that emphasizes only short-term target setting and move to an effective strategic approach. It examines early evidence from research on what dimensions are critical in establishing a strategically focused school within a medium-term…

  15. Strategic Planning--Definition, Process, and Outcomes.

    ERIC Educational Resources Information Center

    Stewart, G. Kent; Bailey, Gerald D.

    1991-01-01

    Strategic planning provides a comprehensive, systematic, and flexible approach to charting direction. Lists an eight-step process and questions for strategic planning. Emphasizes the role of architects in strategic planning. (five references) (MLF)

  16. A Cre-inducible fluorescent reporter for observing apical membrane dynamics.

    PubMed

    Pan, Xinchao; Schnell, Ulrike; Karner, Courtney M; Small, Erin V; Carroll, Thomas J

    2015-01-01

    The ability to image living tissues with fluorescent proteins has revolutionized the fields of cell and developmental biology. Fusions between fluorescent proteins and various polypeptides are allowing scientists to image tissues with sub-cellular resolution. Here, we describe the generation and activity of a genetically engineered mouse line expressing a fusion between the green fluorescent protein (GFP) and the apically localized protein Crumbs3 (Crb3). This reporter drives Cre-inducible expression of Crb3-GFP under control of the EF1a regulatory domains. The fusion protein is broadly expressed in embryonic and adult tissues and shows apical restriction in the majority of epithelial cell types. It displays a variably penetrant gain of function activity in the neural tube. However, in several cell types, over-expression of Crb3 does not appear to have any effect on normal development or maintenance. Detailed analysis of kidneys expressing this reporter indicates normal morphology and function highlighting the utility for live imaging. Thus, the EF1a(Crb3-GFP) mouse line will be of broad use for studying membrane and/or tissue dynamics in living tissues. PMID:25809849

  17. Cylindrically symmetric wormholes WhCRe : The motion of test particles

    NASA Astrophysics Data System (ADS)

    Aminova, Asya V.; Brill, Dieter R.; Chumarov, Pavel I.; Shemakhin, Aleksandr Yu.

    2016-07-01

    We study the radial and nonradial motion of massive test particles and photons in a three-parameter family of cylindrically symmetric wormholes WhCRe generated by the electromagnetic, dilaton, and scalar fields, with particular attention paid to the extent to which the wormhole is traversable. The wormholes are not asymptotically flat and contain a curvature singularity at one end of the wormhole. In the case of nonradial motion with conserved energy and angular momentum along a hypersurface z =const ("planar orbits") we show that, as in the Kerr and Schwarzschild geometries, we should distinguish between orbits with impact parameters greater or less than a certain critical value Dc, which corresponds to the unstable circular orbit of radius uc. For D2>Dc2 there are two kinds of orbits: orbits of the first kind arrive from infinity and turn around at the orbit's minimum radial coordinate u ("pericenter") greater than uc, whereas orbits of the second kind turn around at maximum radial coordinate u ("apocenter") less than uc and terminate at the singularity at u =-∞ . For D =Dc orbits of the first and second kinds merge and both orbits spiral an infinite number of times toward the unstable circular orbit u =uc. For D2

  18. The conditional KO approach: Cre/Lox technology in human neurons

    PubMed Central

    Patzke, Christopher; Südhof, Thomas C.

    2016-01-01

    ABSTRACT The use of human pluripotent stem cells to model human diseases has become a new standard in biomedical sciences. To this end, patient-derived somatic cells are studied in vitro to mimic human pathological conditions. Here, we describe an alternative experimental strategy, the ‘conditional KO approach’, which allows engineering disease-relevant mutations in pluripotent stem cells from healthy donors. In combination with the Cre/Lox technology, this strategy enables us to study the molecular causes of human diseases independent of the genetic background or of genetic alterations induced by clonal selection. As a proof-of-principle, we generated pluripotent stem cells with conditional loss-of-function mutations in the human STXBP1 gene that encodes Munc18-1. Using neurons derived from these cells, we show that heterozygous disruption of STXBP1 produces a specific and selective impairment in synaptic transmission that may account for the severe neurological disease caused by such mutations in human patients. PMID:27141410

  19. Genetic Manipulations Reveal Dynamic Cell and Gene Functions: Cre-ating a New View of Myogenesis

    PubMed Central

    Hutcheson, David A.; Kardon, Gabrielle

    2010-01-01

    Development of multicellular organisms is temporally and spatially complex. The Cre/loxP and Flp/FRT systems for genetic manipulation in mammals now enable researchers to explicitly examine in vivo the temporal and spatial role of cells and genes during development via cell lineage and ablation studies and conditional gene inactivation and activation. Recently we have used these methods to genetically dissect the role of Pax3+ and Pax7+ progenitor populations and the function of β-catenin, an important regulator of myogenesis, in vertebrate limb myogenesis. Our lineage and ablation studies of Pax3+ and Pax7+ progenitors revealed surprising insights into myogenesis not apparent from Pax3 and Pax7 expression and functional studies. In addition, conditional inactivation and activation of β-catenin in different progenitor populations and their progeny demonstrated that β-catenin plays several cell-autonomous roles in myogenesis. Our studies highlight the hierarchical (i.e. genes versus cells), temporal, and spatial complexity of development and demonstrate that manipulations of both cells and genes will be required to obtain a full understanding of the development of multicellular organisms. PMID:19844163

  20. Camk2a-Cre-mediated conditional deletion of chromatin remodeler Brg1 causes perinatal hydrocephalus.

    PubMed

    Cao, Mou; Wu, Jiang I

    2015-06-15

    Mammalian SWI/SNF-like BAF chromatin remodeling complexes are essential for many aspects of neural development. Mutations in the genes encoding the core subunit Brg1/SmarcA4 or other complex components cause neurodevelopmental diseases and are associated with autism. Congenital hydrocephalus is a serious brain disorder often experienced by these patients. We report a role of Brg1 in the pathogenesis of hydrocephalus disorder. We discovered an unexpected early activity of mouse Camk2a-Cre transgene, which mediates Brg1 deletion in a subset of forebrain neurons beginning in the late embryonic stage. Brg1 deletion in these neurons led to severe congenital hydrocephalus with enlargement of the lateral ventricles and attenuation of the cerebral cortex. The Brg1-deficient mice had significantly smaller subcommissural organs and narrower Sylvian aqueducts than mice that express normal levels of Brg1. Effects were non-cell autonomous and may be responsible for the development of the congenital hydrocephalus phenotype. Our study provides evidence indicating that abnormalities in Brg1 function result in defects associated with neurodevelopmental disorders and autism. PMID:25929186

  1. Visualizing the lateral somitic frontier in the Prx1Cre transgenic mouse

    PubMed Central

    Durland, J Logan; Sferlazzo, Matteo; Logan, Malcolm; Burke, Ann Campbell

    2008-01-01

    Changes in the organization of the musculoskeletal system have accounted for many evolutionary adaptations in the vertebrate body plan. The musculoskeletal system develops from two mesodermal populations: somitic mesoderm gives rise to the axial skeleton and all of the skeletal muscle of the body, and lateral plate mesoderm gives rise to the appendicular skeleton. The recognition of embryonic domains resulting from the dynamics of morphogenesis has inspired new terminology based on developmental criteria. Two mesodermal domains are defined, primaxial and abaxial. The primaxial domain includes musculoskeletal structures comprising just somitic cells. The abaxial domain contains somitic myoblasts in connective tissue derived from lateral plate mesoderm, as well as lateral plate-derived skeletal structures. The boundary between these two domains is the lateral somitic frontier. Recent studies have described the developmental relationship between these two domains in the chick. In the present study, we describe the labelling pattern in the body of the Prx1/Cre/Z/AP compound transgenic mouse. The enhancer employed in this transgenic leads to reporter expression in the postcranial, somatic lateral plate mesoderm. The boundary between labelled and unlabelled cell populations is described at embryonic day (E)13.5 and E15.5. We argue that the distribution of labelled cells is consistent with the somatic lateral plate lineage, and therefore provides an estimate of the position of the lateral somitic frontier. The role of the frontier in both development and evolution is discussed. PMID:18430087

  2. A CaMK cascade activates CRE-mediated transcription in neurons of Caenorhabditis elegans

    PubMed Central

    Kimura, Yoshishige; Corcoran, Ethan E.; Eto, Koh; Gengyo-Ando, Keiko; Muramatsu, Masa-aki; Kobayashi, Ryoji; Freedman, Jonathan H.; Mitani, Shohei; Hagiwara, Masatoshi; Means, Anthony R.; Tokumitsu, Hiroshi

    2002-01-01

    Calcium (Ca2+) signals regulate a diverse set of cellular responses, from proliferation to muscular contraction and neuro-endocrine secretion. The ubiquitous Ca2+ sensor, calmodulin (CaM), translates changes in local intracellular Ca2+ concentrations into changes in enzyme activities. Among its targets, the Ca2+/CaM-dependent protein kinases I and IV (CaMKs) are capable of transducing intraneuronal signals, and these kinases are implicated in neuronal gene regulation that mediates synaptic plasticity in mammals. Recently, the cyclic AMP response element binding protein (CREB) has been proposed as a target for a CaMK cascade involving not only CaMKI or CaMKIV, but also an upstream kinase kinase that is also CaM regulated (CaMKK). Here, we report that all components of this pathway are coexpressed in head neurons of Caenorhabditis elegans. Utilizing a transgenic approach to visualize CREB-dependent transcription in vivo, we show that this CaMK cascade regulates CRE-mediated transcription in a subset of head neurons in living nematodes. PMID:12231504

  3. FLP/FRT and Cre/lox recombination technology in C. elegans

    PubMed Central

    2014-01-01

    One of the most powerful aspects of biological inquiry using model organisms is the ability to control gene expression. A holy grail is both temporal and spatial control of the expression of specific gene products – that is, the ability to express or withhold the activity of genes or their products in specific cells at specific times. Ideally such a method would also regulate the precise levels of gene activity, and alterations would be reversible. The related goal of controlled or purposefully randomized expression of visible markers is also tremendously powerful. While not all of these feats have been accomplished in C. elegans to date, much progress has been made, and recent technologies put these goals within closer reach. Here, I present published examples of successful two-component site-specific recombination in C. elegans. These technologies are based on the principle of controlled intra-molecular excision or inversion of DNA sequences between defined sites, as driven by FLP or Cre recombinases. I discuss several prospects for future applications of this technology. PMID:24874786

  4. Flow cytometry and single nucleus sorting for Cre-based analysis of changes in transcriptional states.

    PubMed

    Samadder, Partha; Weng, Ning; Doetschman, Thomas; Heimark, Ronald L; Galbraith, David W

    2016-05-01

    The organs of eukaryotic organisms comprise complex interspersions of cell types, whose different molecular activities, and corresponding cellular states, cooperate during development to produce the final, functional organ. Dysfunction of organs in disease, particularly oncogenesis, initiates with changes of state of a minor subset of cells. It therefore is hard to detect early molecular indicators of disease within an overwhelming background of normal cells. Flow cytometry and sorting provides a convenient way to purify minority subpopulations, if a specific fluorophore can be unambiguously and exclusively associated with this subpopulation. We have generated a number of transgenic mouse lines expressing a nuclear-localized version of the Green Fluorescent Protein (GFP), within which the production of a chimeric histone 2B-GFP protein occurs under the control of a constitutively-active, actin-derived promoter, separated by a Floxed-STOP sequence. In the presence of Cre recombinase, within F1 progeny of these mouse lines, excision of the STOP sequence activates transcription which results in the emergence of cells containing green fluorescent nuclei. We describe the characterization of these lines using a combination of microscopic imaging, flow cytometry and sorting, and Reverse-Transcription polymerase chain reaction of transcripts within single sorted nuclei isolated from tissue homogenates. These lines should be particularly useful for analysis of transcriptional changes in oncogenesis. © 2016 International Society for Advancement of Cytometry. PMID:27003621

  5. Strategic cycling: shaking complacency in healthcare strategic planning.

    PubMed

    Begun, J; Heatwole, K B

    1999-01-01

    As the conditions affecting business and healthcare organizations in the United States have become more turbulent and uncertain, strategic planning has decreased in popularity. Strategic planning is criticized for stiffling creative responses to the new marketplace and for fostering compartmentalized organizations, adherence to outmoded strategies, tunnel vision in strategy formulation, and overemphasis on planning to the detriment of implementation. However, effective strategic planning can be a force for mobilizing all the constituents of an organization, creating discipline in pursuit of a goal, broadening an organization's perspective, improving communication among disciplines, and motivating the organization's workforce. It is worthwhile for healthcare organizations to preserve these benefits of strategic planning at the same time recognizing the many sources of turbulence and uncertainty in the healthcare environment. A model of "strategic cycling" is presented to address the perceived shortcomings of traditional strategic planning in a dynamic environment. The cycling model facilitates continuous assessment of the organization's mission/values/vision and primary strategies based on feedback from benchmark analysis, shareholder impact, and progress in strategy implementation. Multiple scenarios and contingency plans are developed in recognition of the uncertain future. The model represents a compromise between abandoning strategic planning and the traditional, linear model of planning based on progress through predetermined stages to a masterpiece plan. PMID:10621138

  6. 2006 NASA Strategic Plan

    NASA Technical Reports Server (NTRS)

    2006-01-01

    On January 14, 2004, President George W. Bush announced A Renewed Spirit of Discovery: The President's Vision for U.S. Space Exploration, a new directive for the Nation's space program. The fundamental goal of this directive is "to advance U.S. scientific, security, and economic interests through a robust space exploration program." In issuing it, the President committed the Nation to a journey of exploring the solar system and beyond: returning to the Moon in the next decade, then venturing further into the solar system, ultimately sending humans to Mars and beyond. He challenged NASA to establish new and innovative programs to enhance understanding of the planets, to ask new questions, and to answer questions that are as old as humankind. NASA enthusiastically embraced the challenge of extending a human presence throughout the solar system as the Agency's Vision, and in the NASA Authorization Act of 2005, Congress endorsed the Vision for Space Exploration and provided additional guidance for implementation. NASA is committed to achieving this Vision and to making all changes necessary to ensure success and a smooth transition. These changes will include increasing internal collaboration, leveraging personnel and facilities, developing strong, healthy NASA Centers,a nd fostering a safe environment of respect and open communication for employees at all levels. NASA also will ensure clear accountability and solid program management and reporting practices. Over the next 10 years, NASA will focus on six Strategic Goals to move forward in achieving the Vision for Space Exploration. Each of the six Strategic Goals is clearly defined and supported by multi-year outcomes that will enhance NASA's ability to measure and report Agency accomplishments in this quest.

  7. Cre recombinase-regulated Endothelin1 transgenic mouse lines: novel tools for analysis of embryonic and adult disorders

    PubMed Central

    Tavares, Andre L.P.; Clouthier, David E.

    2015-01-01

    Endothelin-1 (EDN1) influences both craniofacial and cardiovascular development and a number of adult physiological conditions by binding to one or both of the known endothelin receptors, thus initiating multiple signaling cascades. Animal models containing both conventional and conditional loss of the Edn1 gene have been used to dissect EDN1 function in both embryos and adults. However, while transgenic Edn1 over-expression or targeted genomic insertion of Edn1 has been performed to understand how elevated levels of Edn1 result in or exacerbate disease states, an animal model in which Edn1 over-expression can be achieved in a spatiotemporal-specific manner has not been reported. Here we describe the creation of Edn1 conditional over-expression transgenic mouse lines in which the chicken β-actin promoter and an Edn1 cDNA are separated by a strong stop sequence flanked by loxP sites. In the presence of Cre, the stop cassette is removed, leading to Edn1 expression. Using the Wnt1-Cre strain, in which Cre expression is targeted to the Wnt1-expressing domain of the central nervous system (CNS) from which neural crest cells (NCCs) arise, we show that stable CBA-Edn1 transgenic lines with varying EDN1 protein levels develop defects in NCC-derived tissues of the face, though the severity differs between lines. We also show that Edn1 expression can be achieved in other embryonic tissues utilizing other Cre strains, with this expression also resulting in developmental defects. CBA-Edn1 transgenic mice will be useful in investigating diverse aspects of EDN1-mediated-development and disease, including understanding how NCCs achieve and maintain a positional and functional identity and how aberrant EDN1 levels can lead to multiple physiological changes and diseases. PMID:25725491

  8. Generation of Shox2-Cre allele for tissue specific manipulation of genes in the developing heart, palate, and limb.

    PubMed

    Sun, Cheng; Zhang, Tao; Liu, Chao; Gu, Shuping; Chen, YiPing

    2013-07-01

    Shox2 is expressed in several developing organs in a tissue specific manner in both mice and humans, including the heart, palate, limb, and nervous system. To better understand the spatial and temporal expression patterns of Shox2 and to systematically dissect the genetic cascade regulated by Shox2, we created Shox2-LacZ and Shox2-Cre knock-in mouse lines. We show that the Shox2-LacZ allele expresses beta-galactosidase reporter gene in a fashion that recapitulates the endogenous Shox2 expression pattern in developing organs, including the sinoatrial node (SAN), the anterior portion of the palate, and the proximal region of the limb bud. Conditional deletion of Shox2 in mice carrying the Shox2-Cre allele yielded SAN phenotypes that resemble conventional Shox2 knockout mice. Our results indicate that the Shox2-Cre allele offer a useful tool for tissue specific manipulation of genes in a number of developing organs, particularly in the developing SAN. PMID:23620086

  9. A Tmprss2-CreERT2 Knock-In Mouse Model for Cancer Genetic Studies on Prostate and Colon.

    PubMed

    Gao, Dong; Zhan, Yu; Di, Wei; Moore, Amanda R; Sher, Jessica J; Guan, Youxin; Wang, Shangqian; Zhang, Zeda; Murphy, Devan A; Sawyers, Charles L; Chi, Ping; Chen, Yu

    2016-01-01

    Fusion between TMPRSS2 and ERG, placing ERG under the control of the TMPRSS2 promoter, is the most frequent genetic alteration in prostate cancer, present in 40-50% of cases. The fusion event is an early, if not initiating, event in prostate cancer, implicating the TMPRSS2-positive prostate epithelial cell as the cancer cell of origin in fusion-positive prostate cancer. To introduce genetic alterations into Tmprss2-positive cells in mice in a temporal-specific manner, we generated a Tmprss2-CreERT2 knock-in mouse. We found robust tamoxifen-dependent Cre activation in the prostate luminal cells but not basal epithelial cells, as well as epithelial cells of the bladder and gastrointestinal (GI) tract. The knock-in allele on the Tmprss2 locus does not noticeably impact prostate, bladder, or gastrointestinal function. Deletion of Pten in Tmprss2-positive cells of adult mice generated neoplasia only in the prostate, while deletion of Apc in these cells generated neoplasia only in the GI tract. These results suggest that this new Tmprss2-CreERT2 mouse model will be a useful resource for genetic studies on prostate and colon. PMID:27536883

  10. The aging behavior of types 308 and 308CRE stainless steels and its effect on mechanical properties

    SciTech Connect

    Vitek, J.M.; David, S.A.

    1987-01-01

    Aging of 308 and 308CRE SS was studied at 475 to 850/sup 0/C for aging times up to 10,000 hours. Above 550/sup 0/C, aging of 308 steel resulted in precipitation of carbides and the transformation of ferrite to sigma phase or the formation of sigma phase in initially ferrite-free material. The elevated-temperature aging of 308CRE steel resulted in the precipitation of titanium-rich carbides, nitrides, and sulfides, and the transformation of ferrite to sigma phase. The distribution of precipitates was affected by the initial condition of the materials. The elevated-temperature creep properties, and in particular the improved properties of 308CRE, were related to the precipitate distribution. Below 550/sup 0/C, aging of welded type 308 steel, precipitation of G-phase within the ferrite was observed, as well as the decomposition of ferrite into alpha and alpha prime. With the help of a novel mechanical properties microprobe, which was capable of determining the hardness of the minor constituent ferrite phase, the hardness behavior as a function of aging could be related to the microstructures. These results are interpreted in terms of the potential susceptibility of these alloys to 475/sup 0/C embrittlement.

  11. A Tmprss2-CreERT2 Knock-In Mouse Model for Cancer Genetic Studies on Prostate and Colon

    PubMed Central

    Zhan, Yu; Di, Wei; Moore, Amanda R.; Sher, Jessica J.; Guan, Youxin; Wang, Shangqian; Zhang, Zeda; Murphy, Devan A.; Sawyers, Charles L.; Chi, Ping; Chen, Yu

    2016-01-01

    Fusion between TMPRSS2 and ERG, placing ERG under the control of the TMPRSS2 promoter, is the most frequent genetic alteration in prostate cancer, present in 40–50% of cases. The fusion event is an early, if not initiating, event in prostate cancer, implicating the TMPRSS2-positive prostate epithelial cell as the cancer cell of origin in fusion-positive prostate cancer. To introduce genetic alterations into Tmprss2-positive cells in mice in a temporal-specific manner, we generated a Tmprss2-CreERT2 knock-in mouse. We found robust tamoxifen-dependent Cre activation in the prostate luminal cells but not basal epithelial cells, as well as epithelial cells of the bladder and gastrointestinal (GI) tract. The knock-in allele on the Tmprss2 locus does not noticeably impact prostate, bladder, or gastrointestinal function. Deletion of Pten in Tmprss2-positive cells of adult mice generated neoplasia only in the prostate, while deletion of Apc in these cells generated neoplasia only in the GI tract. These results suggest that this new Tmprss2-CreERT2 mouse model will be a useful resource for genetic studies on prostate and colon. PMID:27536883

  12. Properties of a Telomerase-Specific Cre/Lox Switch for Transcriptionally Targeted Cancer Gene Therapy1

    PubMed Central

    Bilsland, Alan E.; Fletcher-Monaghan, Aileen; Keith, W. Nicol

    2005-01-01

    Abstract Telomerase expression represents a good target for cancer gene therapy. The promoters of the core telomerase catalytic [human telomerase reverse transcriptase (hTERT)] and RNA [human telomerase RNA (hTR)] subunits show selective activity in cancer cells but not in normal cells. This property can be harnessed to express therapeutic transgenes in a wide range of cancer cells. Unfortunately, weak hTR and hTERT promoter activities in some cancer cells could limit the target cell range. Therefore, strategies to enhance telomerase-specific gene therapy are of interest. We constructed a Cre/Lox reporter switch coupling telomerase promoter specificity with Cytomegalovirus (CMV) promoter activity, which is generally considered to be constitutively high. In this approach, a telomerase-specific vector expressing Cre recombinase directs excisive recombination on a second vector, removing a transcriptional blockade to CMV-dependent luciferase expression. We tested switch activation in cell lines over a wide range of telomerase promoter activities. However, Cre/Lox–dependent luciferase expression was not enhanced relative to expression using hTR or hTERT promoters directly. Cell-specific differences between telomerase and CMV promoter activities and incomplete sigmoid switch activation were limiting factors. Notably, CMV activity was not always significantly stronger than telomerase promoter activity. Our conclusions provide a general basis for a more rational design of novel recombinase switches in gene therapy. PMID:16331888

  13. Evidence from cosmic ray exposure (CRE) dating for the existence of a pre-Minoan caldera on Santorini, Greece

    NASA Astrophysics Data System (ADS)

    Athanassas, C. D.; Bourlès, D. L.; Braucher, R.; Druitt, T. H.; Nomikou, P.; Léanni, L.

    2016-05-01

    Cosmic ray exposure (CRE) dating was performed on the caldera cliffs of Santorini with the aim of detecting cliff segments predating the Minoan eruption (17th century BCE). The methodology involved the determination of in situ-produced cosmogenic 36Cl concentration in basaltic-to-rhyodacitic whole rocks cropping out in the cliffs. After the samples were processed following the chemical protocol of 36Cl preparation for silicate rocks, 36Cl concentrations were measured by accelerator mass spectrometry (AMS). Important challenges during the implementation procedure were related to large amounts of radiogenic 36Cl, complex modeling of inherited 36Cl, and dominance of the thermal and epithermal (low-energy) neutron capture production pathway. Nevertheless, quantitative assessments on the basis of the contribution of the low-energy neutron capture pathway percent to the total production rate validated the calculated CRE dates. Current CRE ages demonstrate that an ancient caldera existed on pre-Minoan Santorini, occupying at least the northern half of the modern-day caldera.

  14. Strategic Petroleum Reserve quarterly report

    SciTech Connect

    Not Available

    1991-08-15

    This August 15, 1991, Strategic Petroleum Reserve Quarterly Report describes activities related to the site development, oil acquisition, budget and cost of the Reserve during the period April 1, 1991, through June 30, 1991. The Strategic Petroleum Reserve storage facilities development program is proceeding on schedule. The Reserve's capacity is currently 726 million barrels. A total of 5.5 million barrels of new gross cavern volume was developed at Big Hill and Bayou Choctaw during the quarter. There were no crude oil deliveries to the Strategic Petroleum Reserve during the calendar quarter ending June 30, 1991. Acquisition of crude oil for the Reserve has been suspended since August 2, 1990, following the invasion of Kuwait by Iraq. As of June 30, 1991, the Strategic Petroleum Reserve inventory was 568.5 million barrels. The reorganization of the Office of the Strategic Petroleum Reserve became effective June 28, 1991. Under the new organization, the Strategic Petroleum Reserve Project Management Office in Louisiana will report to the Strategic Petroleum Reserve Program Office in Washington rather than the Oak Ridge Field Office in Tennessee. 2 tabs.

  15. A Strategic Action Plan for Advancing Math and Science Education in New Mexico 2007-2010

    ERIC Educational Resources Information Center

    New Mexico Public Education Department, 2007

    2007-01-01

    This Strategic Action Plan for Advancing Math and Science Education is an initial outline of strategies, actions, measures of progress, resources needed, timelines, and responsible parties. The Plan focuses on these three main goals: (1) increasing student interest, participation, and achievement in math and science; (2) raising public support and…

  16. Raising School Attendance: A Case Study of Good Practice in Monitoring and Raising Standards

    ERIC Educational Resources Information Center

    Reid, Ken

    2006-01-01

    Purpose: The purpose of this research is to discuss the issue of managing and reducing school absenteeism and truancy. Design/methodology/approach: The article proposes the development and implementation of some long-term strategic approaches to tackling truancy and other forms of non-attendance from school. This article focuses on the innovative…

  17. Characterization of genetically engineered mouse models carrying Col2a1-cre-induced deletions of Lrp5 and/or Lrp6

    PubMed Central

    Schumacher, Cassie A; Joiner, Danese M; Less, Kennen D; Drewry, Melissa Oosterhouse; Williams, Bart O

    2016-01-01

    Mice carrying Collagen2a1-cre-mediated deletions of Lrp5 and/or Lrp6 were created and characterized. Mice lacking either gene alone were viable and fertile with normal knee morphology. Mice in which both Lrp5 and Lrp6 were conditionally ablated via Collagen2a1-cre-mediated deletion displayed severe defects in skeletal development during embryogenesis. In addition, adult mice carrying Collagen2a1-cre-mediated deletions of Lrp5 and/or Lrp6 displayed low bone mass suggesting that the Collagen2a1-cre transgene was active in cells that subsequently differentiated into osteoblasts. In both embryonic skeletal development and establishment of adult bone mass, Lrp5 and Lrp6 carry out redundant functions. PMID:26962465

  18. The Value of Strategic Partnerships

    SciTech Connect

    Gould, Josh; Narayan, Amit; McNutt, Ty

    2015-02-10

    Strong strategic partnerships can be the difference between those technologies that only achieve success in the lab and those that actually break into the marketplace. Two ARPA-E awardees—AutoGrid and APEI—have forged strategic partnerships that have positioned their technologies to achieve major success in the market. This video features remarks from ARPA-E Technology-to-Market Advisor Josh Gould and interviews with technologists at AutoGrid and APEI, who each tell the story of how their company leveraged relationships with strategic partners to broaden their customer base and bring their technology to life.

  19. Inactivation of Tgfbr2 in Osterix-Cre expressing Dental Mesenchyme Disrupts Molar Root Formation

    PubMed Central

    Coricor, George; MacDougall, Mary; Serra, Rosa

    2013-01-01

    It has been difficult to examine the role of TGF-ß in post-natal tooth development due to perinatal lethality in many of the signaling deficient mouse models. To address the role of Tgfbr2 in postnatal tooth development, we generated a mouse in which Tgfbr2 was deleted in odontoblast-and bone-producing mesenchyme. Osx-Cre;Tgfbr2fl/fl mice were generated (Tgfbr2cko) and postnatal tooth development was compared in Tgfbr2cko and control littermates. X-ray and μCT analysis showed that in Tgfbr2cko mice radicular dentin matrix density was reduced in the molars. Molar shape was abnormal and molar eruption was delayed in the mutant mice. Most significantly, defects in root formation, including failure of the root to elongate, were observed by postnatal day 10. Immunostaining for Keratin-14 (K14) was used to delineate Hertwig's epithelial root sheath (HERS). The results showed a delay in elongation and disorganization of the HERS in Tgfbr2cko mice. In addition, the HERS was maintained and the break up into epithelial rests was attenuated suggesting that Tgfbr2 acts on dental mesenchyme to indirectly regulate the formation and maintenance of the HERS. Altered odontoblast organization and reduced Dspp expression indicated that odontoblast differentiation was disrupted in the mutant mice likely contributing to the defect in root formation. Nevertheless, expression of Nfic, a key mesenchymal regulator of root development, was similar in Tgfbr2cko mice and controls. The number of osteoclasts in the bone surrounding the tooth was reduced and osteoblast differentiation was disrupted likely contributing to both root and eruption defects. We conclude that Tgfbr2 in dental mesenchyme and bone is required for tooth development particularly root formation. PMID:23933490

  20. Carbon catabolite repressor gene BbCre1 influences carbon source uptake but does not have a big impact on virulence in Beauveria bassiana.

    PubMed

    Jin, Kai; Luo, Zhibing; Jiang, Xiaodong; Zhang, Yongjun; Zhou, Yonghong; Pei, Yan

    2011-03-01

    A gene (BbCre1, GenBank accession number EF108309) encoding a carbon catabolite repressor (CreA) with two Cys(2)His(2) zinc finger regions and a nuclear localization signal was cloned from the entomopathogenic fungus Beauveria bassiana. Overexpression and antisense strategies were used to investigate the biological functions of this gene. Compared with the wild type, the conidial yield and colony growth of BbCre1-overexpression transformants were significantly decreased on the plates with xylose or ethanol as the sole carbon source. With glucose as the sole carbon source, a significant difference was observed in the activity of Pr1A-like protease among BbCre1-overexpression transformants, antisense-BbCre1 transformants and the wild type. However, bioassays showed that knockdown or overexpression of BbCre1 did not have a significant impact on the virulence of B. bassiana to aphids. These results imply that the fungus remains virulent, even when simpler, less expensive nutrients are available, i.e. glucose. PMID:21130770

  1. Sox2-CreER mice are useful for fate mapping of mature, but not neonatal, cochlear supporting cells in hair cell regeneration studies

    PubMed Central

    Walters, Bradley J.; Yamashita, Tetsuji; Zuo, Jian

    2015-01-01

    Studies of hair cell regeneration in the postnatal cochlea rely on fate mapping of supporting cells. Here we characterized a Sox2-CreER knock-in mouse line with two independent reporter mouse strains at neonatal and mature ages. Regardless of induction age, reporter expression was robust, with CreER activity being readily detectable in >85% of supporting cells within the organ of Corti. When induced at postnatal day (P) 28, Sox2-CreER activity was exclusive to supporting cells demonstrating its utility for fate mapping studies beyond this age. However, when induced at P1, Sox2-CreER activity was also detected in >50% of cochlear hair cells, suggesting that Sox2-CreER may not be useful to fate map a supporting cell origin of regenerated hair cells if induced at neonatal ages. Given that this model is currently in use by several investigators for fate mapping purposes, and may be adopted by others in the future, our finding that current protocols are effective for restricting CreER activity to supporting cells at mature but not neonatal ages is both significant and timely. PMID:26108463

  2. BTS strategic plan executive summary

    SciTech Connect

    1999-01-06

    This Strategic Plan positions the Office of Building Technology, State and Community Programs (BTS) to be more effective in reducing energy consumption in homes, offices, schools and other buildings, and in reducing environmental impacts associated with energy use.

  3. Financial Management and Strategic Planning.

    ERIC Educational Resources Information Center

    Dickmeyer, Nathan

    1982-01-01

    Strategic planning involves analysis of risks and opportunities; financial management entails balancing risks and resources to achieve institutional goals. A framework is suggested to help college administrators integrate the two functions. (Author/MSE)

  4. 7 CFR 25.202 - Strategic plan.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 1 2011-01-01 2011-01-01 false Strategic plan. 25.202 Section 25.202 Agriculture Office of the Secretary of Agriculture RURAL EMPOWERMENT ZONES AND ENTERPRISE COMMUNITIES Nomination Procedure § 25.202 Strategic plan. (a) Principles of strategic plan. The strategic plan included in the application must be developed in accordance...

  5. 29 CFR 780.123 - Raising of bees.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 3 2012-07-01 2012-07-01 false Raising of bees. 780.123 Section 780.123 Labor Regulations... Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.123 Raising of bees. The term “raising of * * * bees” refers to all of those activities customarily performed in connection with...

  6. 29 CFR 780.123 - Raising of bees.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 3 2011-07-01 2011-07-01 false Raising of bees. 780.123 Section 780.123 Labor Regulations... Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.123 Raising of bees. The term “raising of * * * bees” refers to all of those activities customarily performed in connection with...

  7. 29 CFR 780.123 - Raising of bees.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 29 Labor 3 2014-07-01 2014-07-01 false Raising of bees. 780.123 Section 780.123 Labor Regulations... Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.123 Raising of bees. The term “raising of * * * bees” refers to all of those activities customarily performed in connection with...

  8. 29 CFR 780.123 - Raising of bees.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 3 2010-07-01 2010-07-01 false Raising of bees. 780.123 Section 780.123 Labor Regulations... Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.123 Raising of bees. The term “raising of * * * bees” refers to all of those activities customarily performed in connection with...

  9. 29 CFR 780.123 - Raising of bees.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 29 Labor 3 2013-07-01 2013-07-01 false Raising of bees. 780.123 Section 780.123 Labor Regulations... Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.123 Raising of bees. The term “raising of * * * bees” refers to all of those activities customarily performed in connection with...

  10. Strategic power: USA/USSR

    SciTech Connect

    Jacobsen, C.G. )

    1989-01-01

    This uniquely comprehensive book details the impact on U.S. and Soviet strategic perceptions of different strategic cultures. It compares and contrasts power, their sometimes quite different interpretation of strategic concepts and reality, and the relevance of analogous yet distinct processes (e.g. decision-making in defence) and dynamics (the impact of military - industrial complexes). It also details the ramifications of new thinking East and West. It is designed for policy-makers, media and lay people who wish to understand the similarities and peculiarities of U.S. and Soviet strategic mindsets, and their approach to strategic opportunity, threat and dilemma. The author list reads like a veritable Who's Who of the best and the brightest of U.S., Soviet, British, Australian and Canadian strategic analysts- a truly exceptional assemblage of expertise. Each chapter may be read alone, for succinct coverage of either U.S. or Soviet though, approach or reality on any particular issue; together, they provide an encyclopedic reference source for governments, students and others.

  11. Raising children in America: Korean parents' experiences.

    PubMed

    Choi, H; Dancy, B L; Lee, J

    2013-08-01

    This qualitative description study was designed to describe Korean American parents' perceptions of challenges and difficulties they encounter while raising their children in the USA. A convenience sampling of 21 parents of adolescents aged 11-14 years recruited from the Midwest Korean American community participated in the study. Data were collected using in-depth, face-to-face interviews, which took place in agreed-upon, convenient locations. All interviews were audiotaped and transcribed in Korean and the transcripts were translated into English. Qualitative content analysis revealed that the main stresses that parents encountered while raising their children in the USA were inability to advocate for children, feeling uneasy and insecure about incompatible American culture, ambivalence towards children's ethnic identities, and feeling alienated. In relation to these stresses, parents often felt inadequate, ashamed, guilty, regretful and powerless. The findings demonstrated the importance of understanding parents' feelings that are deeply embedded in the conflicted parent-child relationships and their perceptions of being parents in the USA. The present study highlights the need for and importance of providing intervention programmes for parents, particularly programmes that would empower parents, strengthen parent-child relationships and address ways to integrate two very different cultures while upholding ethnic identity and pride. PMID:22295897

  12. IPPF's strategic vision 2000.

    PubMed

    Mahler, H

    1994-09-01

    The International Planned Parenthood Federation (IPPF) supports the objectives of the International Conference on Population and Development. The draft conference document displays the linkages between women's rights, poverty, population, and sustainable development. In the next two to three decades the global population will grow by a staggering 50% and that reality requires sustained socioeconomic growth. The 6 components of IPPF's Strategic Vision 2000 and the action agenda for meeting these challenges are as follows: 1) The small family is rapidly becoming the cultural norm everywhere, as more than 60% of all fertile women and men are using safe and effective contraceptive methods. 2) Sexual and reproductive health is becoming an integral part of the physical, mental, and social health culture and not just the absence of pregnancy, disease or injury. The members of IPPF have been developing programs concentrating on adolescent sexuality. IPPF is also concerned about HIV, AIDS and sexually transmitted diseases as they relate to the work undertaken by family planning associations. 3) The quality of care has been heightened by the right of informed choice, service sustainability, and the maintenance of the highest standards. 4) Vigorous programs are in place to eliminate unsafe abortion and to increase access to safe abortion. 5) Young people are systematically supported to give voice to the need for understanding their sexuality and services. There are currently more than 500 million young women and men between the ages of 15 and 19, and their numbers increase for a long time to come. 6) Women are fast moving towards full participation in all aspects of local and global development demanding reproductive rights. IPPF and its associations, through Vision 2000, are committed to the objective of sexual and reproductive health at the national and international levels. PMID:12288265

  13. The need for strategic tax planning among nonprofit hospitals.

    PubMed

    Smith, Pamela C

    2005-01-01

    Strategic tax planning issues are important to the nonprofit health care sector, despite its philanthropic mission. The consolidation of the industry has led management to fight for resources and develop alternative strategies for raising money. When management evaluates alternative collaborative structures to increase efficiency, the impact on governance structures must also be considered. The increased governmental scrutiny of joint ventures within the health care sector warrants management's attention as well. The financial incentives must be considered, along with the various tax policy implications of cross-sector collaborations. PMID:18973000

  14. The Pseudomonas aeruginosa CreBC two-component system plays a major role in the response to β-lactams, fitness, biofilm growth, and global regulation.

    PubMed

    Zamorano, Laura; Moyà, Bartolomé; Juan, Carlos; Mulet, Xavier; Blázquez, Jesús; Oliver, Antonio

    2014-09-01

    Pseudomonas aeruginosa is a ubiquitous versatile environmental microorganism with a remarkable ability to grow under diverse environmental conditions. Moreover, P. aeruginosa is responsible for life-threatening infections in immunocompromised and cystic fibrosis patients, as the extraordinary capacity of this pathogen to develop antimicrobial resistance dramatically limits our therapeutic arsenal. Its large genome carries an outstanding number of genes belonging to regulatory systems, including multiple two-component sensor-regulator systems that modulate the response to the different environmental stimuli. Here, we show that one of two systems, designated CreBC (carbon source responsive) and BlrAB (β-lactam resistance), might be of particular relevance. We first identified the stimuli triggering the activation of the CreBC system, which specifically responds to penicillin-binding protein 4 (PBP4) inhibition by certain β-lactam antibiotics. Second, through an analysis of a large comprehensive collection of mutants, we demonstrate an intricate interconnection between the CreBC system, the peptidoglycan recycling pathway, and the expression of the concerning chromosomal β-lactamase AmpC. Third, we show that the CreBC system, and particularly its effector inner membrane protein CreD, plays a major role in bacterial fitness and biofilm development, especially in the presence of subinhibitory concentrations of β-lactams. Finally, global transcriptomics reveals broad regulatory functions of CreBC in basic physiological aspects, particularly anaerobic respiration, in both the presence and absence of antibiotics. Therefore, the CreBC system is envisaged as a potentially interesting target for improving the efficacy of β-lactams against P. aeruginosa infections. PMID:24936599

  15. Targeted expression in zebrafish primordial germ cells by Cre/loxP and Gal4/UAS systems.

    PubMed

    Xiong, Feng; Wei, Zhi-Qiang; Zhu, Zuo-Yan; Sun, Yong-Hua

    2013-10-01

    In zebrafish and other vertebrates, primordial germ cells (PGCs) are a population of embryonic cells that give rise to sperm and eggs in adults. Any type of genetically manipulated lines have to be originated from the germ cells of the manipulated founders, thus it is of great importance to establish an effective technology for highly specific PGC-targeted gene manipulation in vertebrates. In the present study, we used the Cre/loxP recombinase system and Gal4/UAS transcription system for induction and regulation of mRFP (monomer red fluorescent protein) gene expression to achieve highly efficient PGC-targeted gene expression in zebrafish. First, we established two transgenic activator lines, Tg(kop:cre) and Tg(kop:KalTA4), to express the Cre recombinases and the Gal4 activator proteins in PGCs. Second, we generated two transgenic effector lines, Tg(kop:loxP-SV40-loxP-mRFP) and Tg(UAS:mRFP), which intrinsically showed transcriptional silence of mRFP. When Tg(kop:cre) females were crossed with Tg(kop:loxP-SV40-loxP-mRFP) males, the loxP flanked SV40 transcriptional stop sequence was 100 % removed from the germ cells of the transgenic hybrids. This led to massive production of PGC-specific mRFP transgenic line, Tg(kop:loxP-mRFP), from an mRFP silent transgenic line, Tg(kop:loxP-SV40-loxP-mRFP). When Tg(kop:KalTA4) females were crossed with Tg(UAS:mRFP) males, the hybrid embryos showed PGC specifically expressed mRFP from shield stage till 25 days post-fertilization (pf), indicating the high sensitivity, high efficiency, and long-lasting effect of the Gal4/UAS system. Real-time PCR analysis showed that the transcriptional amplification efficiency of the Gal4/UAS system in PGCs can be about 300 times higher than in 1-day-pf embryos. More importantly, when the UAS:mRFP-nos1 construct was directly injected into the Tg(kop:KalTA4) embryos, it was possible to specifically label the PGCs with high sensitivity, efficiency, and persistence. Therefore, we have established two

  16. 1995-1996 Strategic Plan

    SciTech Connect

    1995-09-01

    In a rapidly changing world, effective governmental organizations must anticipate and plan for the future. Nowhere is this more true than in the realm of information generation and dissemination. The Energy Information Administration (EIA) faces a future of great opportunity and also significant challenge. Technological advances are providing opportunities to improve our work processes and the way we serve our customers. This year`s Strategic Plan was developed by an EIA Strategic Planning Group with a membership that spans the entire organization. The Strategic Plan provides both a broad vision and a clear map of EIA`s near-term future. The Strategic Planning Group conducted an analysis of the strengths and weaknesses of EIA as an organization, and of the opportunities and constraints facing EIA in the future. Opportunities exist in the future in technological advances and staff training; constraints lie in federal budget trends and staff demographics. This will require a Strategic Plan that recognizes the need to prioritize services and products across EIA, standardize technology across EIA where possible, lead with fewer layers of management, and be acutely aware of customer needs. Developing and training EIA employees to meet the challenges of rapidly changing technology and decreasing budgets were given high priority.

  17. Fund-Raising While You Shop: A Look at Fund-Raising Sites on the Internet.

    ERIC Educational Resources Information Center

    Seydel, Angela

    2000-01-01

    Explains how school fund-raising can be done over the Internet through electronic commerce. Discusses online retailers who donate percentage of sales to schools who have registered; Web sites that coordinate online retailers for schools, who also take a percentage of the sales; loss of revenue for local businesses; and marketing. (LRW)

  18. Strategic Petroleum Reserve quarterly report

    SciTech Connect

    1995-11-15

    The Strategic Petroleum Reserve was created pursuant to the Energy Policy and Conservation Act of December 22, 1975 (Public Law 94-163). Its purposes are to reduce the impact of disruptions in supplies of petroleum products and to carry out obligations of the United States under the Agreement on an International Energy Program. Section 165(a) of the Act requires the submission of Annual Reports and Section 165(b)(1) requires the submission of Quarterly Reports. This Quarterly Report highlights activities undertaken during the third quarter of calendar year 1995, including: inventory of petroleum products stored in the Reserve; current storage capacity and ullage available; current status of the Strategic Petroleum Reserve storage facilities, major projects and the acquisition of petroleum products; funds obligated by the Secretary from the SPR Petroleum Account and the Strategic Petroleum Reserve Account during the prior calendar quarter and in total; and major environmental actions completed, in progress, or anticipated.

  19. Strategic Asia 2002 Final Report

    SciTech Connect

    Richard Ellings; Aaron Friedberg; Michael Wills

    2002-09-01

    The Strategic Asia Program made considerable progress over the course of 2002--the program's first year with support from the Department of Energy--and completed all its tasks on schedule and within budget. Following a planning meeting in Washington in February 2002, a team of leading specialists wrote a series of original assessments regarding the impact of September 11 on the strategic environment in Asia, examining how perceptions and strategies of countries in the region changed following the terrorist attacks. The final products, Strategic Asia 2002-03: Asian Aftershocks and its accompanying executive summary, were published in September 2002. The program's research findings (some of which are summarized) were presented to policymakers in Washington and elsewhere throughout the year, and almost 2,000 copies of the book had been distributed by mid-2003.

  20. Generation of brain tumours in mice by Cre-mediated recombination of neural progenitors in situ with the tamoxifen metabolite endoxifen.

    PubMed

    Benedykcinska, Anna; Ferreira, Andreia; Lau, Joanne; Broni, Jessica; Richard-Loendt, Angela; Henriquez, Nico V; Brandner, Sebastian

    2016-02-01

    Targeted cell- or region-specific gene recombination is widely used in the functional analysis of genes implicated in development and disease. In the brain, targeted gene recombination has become a mainstream approach to study neurodegeneration or tumorigenesis. The use of the Cre-loxP system to study tumorigenesis in the adult central nervous system (CNS) can be limited, when the promoter (such as GFAP) is also transiently expressed during development, which can result in the recombination of progenies of different lineages. Engineering of transgenic mice expressing Cre recombinase fused to a mutant of the human oestrogen receptor (ER) allows the circumvention of transient developmental Cre expression by inducing recombination in the adult organism. The recombination of loxP sequences occurs only in the presence of tamoxifen. Systemic administration of tamoxifen can, however, exhibit toxicity and might also recombine unwanted cell populations if the promoter driving Cre expression is active at the time of tamoxifen administration. Here, we report that a single site-specific injection of an active derivative of tamoxifen successfully activates Cre recombinase and selectively recombines tumour suppressor genes in neural progenitor cells of the subventricular zone in mice, and we demonstrate its application in a model for the generation of intrinsic brain tumours. PMID:26704996

  1. Generation of brain tumours in mice by Cre-mediated recombination of neural progenitors in situ with the tamoxifen metabolite endoxifen

    PubMed Central

    Benedykcinska, Anna; Ferreira, Andreia; Lau, Joanne; Broni, Jessica; Richard-Loendt, Angela; Henriquez, Nico V.; Brandner, Sebastian

    2016-01-01

    ABSTRACT Targeted cell- or region-specific gene recombination is widely used in the functional analysis of genes implicated in development and disease. In the brain, targeted gene recombination has become a mainstream approach to study neurodegeneration or tumorigenesis. The use of the Cre-loxP system to study tumorigenesis in the adult central nervous system (CNS) can be limited, when the promoter (such as GFAP) is also transiently expressed during development, which can result in the recombination of progenies of different lineages. Engineering of transgenic mice expressing Cre recombinase fused to a mutant of the human oestrogen receptor (ER) allows the circumvention of transient developmental Cre expression by inducing recombination in the adult organism. The recombination of loxP sequences occurs only in the presence of tamoxifen. Systemic administration of tamoxifen can, however, exhibit toxicity and might also recombine unwanted cell populations if the promoter driving Cre expression is active at the time of tamoxifen administration. Here, we report that a single site-specific injection of an active derivative of tamoxifen successfully activates Cre recombinase and selectively recombines tumour suppressor genes in neural progenitor cells of the subventricular zone in mice, and we demonstrate its application in a model for the generation of intrinsic brain tumours. PMID:26704996

  2. Final Draft Strategic Marketing Plan.

    SciTech Connect

    United States. Bonneville Power Administration.

    1994-02-01

    The Bonneville Power Administration (BPA) has developed a marketing plan to define how BPA can be viable and competitive in the future, a result important to BPA`s customers and constituents. The Marketing Plan represents the preferred customer outcomes, marketplace achievements, and competitive advantage required to accomplish the Vision and the Strategic Business Objectives of the agency. The Marketing Plan contributes to successful implementation of BPA`s Strategic Business Objectives (SBOs) by providing common guidance to organizations and activities throughout the agency responsible for (1) planning, constructing, operating, and maintaining the Federal Columbia River Power System; (2) conducting business with BPA`s customers; and (3) providing required internal support services.

  3. Strategic materials availability and supply

    NASA Technical Reports Server (NTRS)

    Stephens, Joseph R.

    1989-01-01

    An evaluation is made of the supply sources and current availability of elements involved in superalloy compositions, as well as of development trends in the substitution of strategic materials for superalloys, giving attention to the results thus far achieved by NASA's Conservation of Strategic Aerospace Materials program, which formally operated only from 1980 to 1984 but whose various research initiatives have been continued under other NASA programs. Peacetime and wartime economies' need and probable availability analyses are presented for bauxite/aluminum, the Pt-group metals, and the alloying elements Mn, Nb, Ta, Co, Cr, Ni, W, Fe, Cu, Ti, Mo, and Mg.

  4. The Strategic Data Project's Strategic Performance Indicators

    ERIC Educational Resources Information Center

    Page, Lindsay C.; Fullerton, Jon; Bacher-Hicks, Andrew; Owens, Antoniya; Cohodes, Sarah R.; West, Martin R.; Glover, Sarah

    2013-01-01

    Strategic Performance Indicators (SPIs) are summary measures derived from parallel, descriptive analyses conducted across educational agencies. The SPIs are designed to inform agency management and efforts to improve student outcomes. We developed the SPIs to reveal patterns common across partner agencies, to highlight exceptions to those…

  5. Strategic Planning and Strategic Thinking Clothed in STRATEGO

    ERIC Educational Resources Information Center

    Baaki, John; Moseley, James L.

    2011-01-01

    This article shares experiences that participants had playing the game of STRATEGO and how the activity may be linked to strategic planning and thinking. Among the human performance technology implications of playing this game are that gamers agreed on a framework for rules, took stock on where they wanted to go in the future, and generated a risk…

  6. Impact of tamoxifen on adipocyte lineage tracing: Inducer of adipogenesis and prolonged nuclear translocation of Cre recombinase

    PubMed Central

    Ye, Risheng; Wang, Qiong A.; Tao, Caroline; Vishvanath, Lavanya; Shao, Mengle; McDonald, Jeffery G.; Gupta, Rana K.; Scherer, Philipp E.

    2015-01-01

    Background The selective estrogen receptor modulator tamoxifen, in combination with the Cre-ERT2 fusion protein, has been one of the mainstream methods to induce genetic recombination and has found widespread application in lineage tracing studies. Methods & results Here, we report that tamoxifen exposure at widely used concentrations remains detectable by mass-spectrometric analysis in adipose tissue after a washout period of 10 days. Surprisingly, its ability to maintain nuclear translocation of the Cre-ERT2 protein is preserved beyond 2 months of washout. Tamoxifen treatment acutely leads to transient lipoatrophy, followed by de novo adipogenesis that reconstitutes the original fat mass. In addition, we find a “synthetically lethal” phenotype for adipocytes when tamoxifen treatment is combined with adipocyte-specific loss-of-function mutants, such as an adipocyte-specific PPARγ knockout. This is observed to a lesser extent when alternative inducible approaches are employed. Conclusions These findings highlight the potential for tamoxifen-induced adipogenesis, and the associated drawbacks of the use of tamoxifen in lineage tracing studies, explaining the discrepancy in lineage tracing results from different systems with temporal control of gene targeting. PMID:26629402

  7. Context dependence between subdomains in the DNA binding interface of the I-CreI homing endonuclease

    PubMed Central

    Grizot, Sylvestre; Duclert, Aymeric; Thomas, Séverine; Duchateau, Philippe; Pâques, Frédéric

    2011-01-01

    Homing endonucleases (HE) have emerged as precise tools for achieving gene targeting events. Redesigned HEs with tailored specificities can be used to cleave new sequences, thereby considerably expanding the number of targetable genes and loci. With HEs, as well as with other protein scaffolds, context dependence of DNA/protein interaction patterns remains one of the major limitations for rational engineering of new DNA binders. Previous studies have shown strong crosstalk between different residues and regions of the DNA binding interface. To investigate this phenomenon, we systematically combined mutations from three groups of amino acids in the DNA binding regions of the I-CreI HE. Our results confirm that important crosstalk occurs throughout this interface in I-CreI. Detailed analysis of success rates identified a nearest-neighbour effect, with a more pronounced level of dependence between adjacent regions. Taken together, these data suggest that combinatorial engineering does not necessarily require the identification of separable functional or structural regions, and that groups of amino acids provide acceptable building blocks that can be assembled, overcoming the context dependency of the DNA binding interface. Furthermore, the present work describes a sequential method to engineer tailored HEs, wherein three contiguous regions are individually mutated and assembled to create HEs with engineered specificity. PMID:21482539

  8. A CRE-DEPENDENT, ANTEROGRADE TRANS-SYNAPTIC VIRAL TRACER FOR MAPPING OUTPUT PATHWAYS OF GENETICALLY MARKED NEURONS

    PubMed Central

    Lo, Liching; Anderson, David J.

    2012-01-01

    SUMMARY Neurotropic viruses that conditionally infect or replicate in molecularly defined neuronal subpopulations, and then spread trans-synaptically, are powerful tools for mapping neural pathways. Genetically targetable retrograde trans-synaptic tracer viruses are available to map the inputs to specific neuronal subpopulations, but an analogous tool for mapping synaptic outputs is not yet available. Here we describe a Cre recombinase-dependent, anterograde trans-neuronal tracer, based on the H129 strain of herpes simplex virus (HSV). Application of this virus to transgenic or knock-in mice expressing Cre in peripheral neurons of the olfactory epithelium or the retina reveals widespread, polysynaptic labeling of higher-order neurons in the olfactory and visual systems, respectively. Polysynaptic pathways were also labeled from cerebellar Purkinje cells. In each system, the pattern of labeling was consistent with classical circuit-tracing studies, restricted to neurons and anterograde-specific. These data provide proof-of-principle for a conditional, non-diluting anterograde trans-synaptic tracer for mapping synaptic outputs from genetically marked neuronal subpopulations. PMID:22196330

  9. Raising the visibility of school nursing services.

    PubMed

    Sanders, Laura; Pratten, Charlie; Pretty, Laura; Roberts, Sheila; Robinson, Jenny

    2016-06-01

    In an attempt to raise the visibility and profile of the school nursing service in the London boroughs of Hounslow and Richmond, school nurses sought to hear from the voices of the young people with whom they work. Ten focus groups in secondary educational provisions were held in a bid to look at young people's accessibility to technological devices, where they would go if they had a health problem and which health topics they would like more information on. This paper examines both national and local data and demographics, and discusses findings from the focus groups run with local young people. The results collected from the focus groups emphasised that technology is a fundamental aspect of many young people's lives and that a large number of young people access the internet to source information about their personal health and wellbeing. The data suggests that the school nursing service could use technologies, such as an interactive app, to support the physical and emotional health of young people. PMID:27443030

  10. Role of the Academic Vice President in Fund-Raising.

    ERIC Educational Resources Information Center

    Felicetti, Daniel A.

    This study explores higher education's failure to incorporate the role of the Academic Vice President (AVP) into campus fund-raising efforts. A literature review found that the Academic Vice President/Provost was rarely mentioned by position as having an instrumental role in fund-raising activities. In a survey of 32 fund-raising consulting firm…

  11. 7 CFR 60.106 - Farm-raised fish.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 3 2011-01-01 2011-01-01 false Farm-raised fish. 60.106 Section 60.106 Agriculture... FOR FISH AND SHELLFISH General Provisions Definitions § 60.106 Farm-raised fish. Farm-raised fish means fish or shellfish that have been harvested in controlled environments, including ocean-ranched...

  12. 29 CFR 780.126 - Contract arrangements for raising poultry.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 3 2011-07-01 2011-07-01 false Contract arrangements for raising poultry. 780.126 Section... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.126 Contract arrangements for raising poultry. Feed dealers and processors sometimes enter into...

  13. 29 CFR 780.125 - Raising of poultry in general.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 29 Labor 3 2013-07-01 2013-07-01 false Raising of poultry in general. 780.125 Section 780.125... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.125 Raising of poultry in general. (a) The term “poultry” includes domesticated fowl and game birds. Ducks...

  14. 29 CFR 780.126 - Contract arrangements for raising poultry.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 29 Labor 3 2013-07-01 2013-07-01 false Contract arrangements for raising poultry. 780.126 Section... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.126 Contract arrangements for raising poultry. Feed dealers and processors sometimes enter into...

  15. 29 CFR 780.125 - Raising of poultry in general.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 3 2012-07-01 2012-07-01 false Raising of poultry in general. 780.125 Section 780.125... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.125 Raising of poultry in general. (a) The term “poultry” includes domesticated fowl and game birds. Ducks...

  16. 29 CFR 780.126 - Contract arrangements for raising poultry.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 3 2012-07-01 2012-07-01 false Contract arrangements for raising poultry. 780.126 Section... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.126 Contract arrangements for raising poultry. Feed dealers and processors sometimes enter into...

  17. 29 CFR 780.125 - Raising of poultry in general.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 29 Labor 3 2014-07-01 2014-07-01 false Raising of poultry in general. 780.125 Section 780.125... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.125 Raising of poultry in general. (a) The term “poultry” includes domesticated fowl and game birds. Ducks...

  18. 29 CFR 780.126 - Contract arrangements for raising poultry.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 29 Labor 3 2014-07-01 2014-07-01 false Contract arrangements for raising poultry. 780.126 Section... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.126 Contract arrangements for raising poultry. Feed dealers and processors sometimes enter into...

  19. 29 CFR 780.126 - Contract arrangements for raising poultry.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 3 2010-07-01 2010-07-01 false Contract arrangements for raising poultry. 780.126 Section... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.126 Contract arrangements for raising poultry. Feed dealers and processors sometimes enter into...

  20. 29 CFR 780.125 - Raising of poultry in general.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 3 2010-07-01 2010-07-01 false Raising of poultry in general. 780.125 Section 780.125... General Scope of Agriculture Raising of Livestock, Bees, Fur-Bearing Animals, Or Poultry § 780.125 Raising of poultry in general. (a) The term “poultry” includes domesticated fowl and game birds. Ducks...

  1. Effective Fund-Raising for Non-profit Camps.

    ERIC Educational Resources Information Center

    Larson, Paula

    1998-01-01

    Identifies and describes strategies for effective fundraising: imagining the possibilities, identifying fund-raising sources, targeting fund-raising efforts, maximizing time by utilizing public relations efforts and involving staff, writing quality proposals and requests, and staying educated on fund-raising topics. Sidebars describe planned…

  2. Verb-raising and Numeral Classifiers in Japanese: Incompatible Bedfellows.

    ERIC Educational Resources Information Center

    Fukushima, Kazuhiko

    2003-01-01

    Examines verb raising in Japanese and looks at Koizumi's (2000) evidence for verb-raising based on data involving, among other things, numeral classifiers. Demonstrates that Koizumi's evidence based on numeral classifiers does not support his claim that verb-raising occurs in Japanese. (Author/VWL)

  3. 5 CFR 2421.21 - Affected by issues raised.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Affected by issues raised. 2421.21 Section 2421.21 Administrative Personnel FEDERAL LABOR RELATIONS AUTHORITY, GENERAL COUNSEL OF THE FEDERAL... § 2421.21 Affected by issues raised. The phrase affected by issues raised, as used in part 2422,...

  4. 7 CFR 60.106 - Farm-raised fish.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 3 2014-01-01 2014-01-01 false Farm-raised fish. 60.106 Section 60.106 Agriculture... FOR FISH AND SHELLFISH General Provisions Definitions § 60.106 Farm-raised fish. Farm-raised fish means fish or shellfish that have been harvested in controlled environments, including ocean-ranched...

  5. 7 CFR 60.106 - Farm-raised fish.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 3 2012-01-01 2012-01-01 false Farm-raised fish. 60.106 Section 60.106 Agriculture... FOR FISH AND SHELLFISH General Provisions Definitions § 60.106 Farm-raised fish. Farm-raised fish means fish or shellfish that have been harvested in controlled environments, including ocean-ranched...

  6. 7 CFR 60.106 - Farm-raised fish.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 3 2013-01-01 2013-01-01 false Farm-raised fish. 60.106 Section 60.106 Agriculture... FOR FISH AND SHELLFISH General Provisions Definitions § 60.106 Farm-raised fish. Farm-raised fish means fish or shellfish that have been harvested in controlled environments, including ocean-ranched...

  7. Divide and conquer: strategic decision areas

    PubMed Central

    Kolling, Nils; Hunt, Laurence T

    2016-01-01

    Human decisions are often strategic, but this can prove difficult to study experimentally. The board game shogi is used to investigate the functional neuroanatomy of strategic decisions, revealing different brain areas to other forms of choice. PMID:25919959

  8. Cre/loxP plus BAC”: a strategy for direct cloning of large DNA fragment and its applications in Photorhabdus luminescens and Agrobacterium tumefaciens

    PubMed Central

    Hu, Shengbiao; Liu, Zhengqiang; Zhang, Xu; Zhang, Guoyong; Xie, Yali; Ding, Xuezhi; Mo, Xiangtao; Stewart, A. Francis; Fu, Jun; Zhang, Youming; Xia, Liqiu

    2016-01-01

    Heterologous expression has been proven to be a valid strategy for elucidating the natural products produced by gene clusters uncovered by genome sequencing projects. Efforts have been made to efficiently clone gene clusters directly from genomic DNA and several approaches have been developed. Here, we present an alternative strategy based on the site-specific recombinase system Cre/loxP for direct cloning gene clusters. A type three secretion system (T3SS) gene cluster (~32 kb) from Photorhabdus luminescens TT01 and DNA fragment (~78 kb) containing the siderophore biosynthetic gene cluster from Agrobacterium tumefaciens C58 have been successfully cloned into pBeloBAC11 with “Cre/loxP plus BAC” strategy. Based on the fact that Cre/loxP system has successfully used for genomic engineering in a wide range of organisms, we believe that this strategy could be widely used for direct cloning of large DNA fragment. PMID:27364376

  9. "Cre/loxP plus BAC": a strategy for direct cloning of large DNA fragment and its applications in Photorhabdus luminescens and Agrobacterium tumefaciens.

    PubMed

    Hu, Shengbiao; Liu, Zhengqiang; Zhang, Xu; Zhang, Guoyong; Xie, Yali; Ding, Xuezhi; Mo, Xiangtao; Stewart, A Francis; Fu, Jun; Zhang, Youming; Xia, Liqiu

    2016-01-01

    Heterologous expression has been proven to be a valid strategy for elucidating the natural products produced by gene clusters uncovered by genome sequencing projects. Efforts have been made to efficiently clone gene clusters directly from genomic DNA and several approaches have been developed. Here, we present an alternative strategy based on the site-specific recombinase system Cre/loxP for direct cloning gene clusters. A type three secretion system (T3SS) gene cluster (~32 kb) from Photorhabdus luminescens TT01 and DNA fragment (~78 kb) containing the siderophore biosynthetic gene cluster from Agrobacterium tumefaciens C58 have been successfully cloned into pBeloBAC11 with "Cre/loxP plus BAC" strategy. Based on the fact that Cre/loxP system has successfully used for genomic engineering in a wide range of organisms, we believe that this strategy could be widely used for direct cloning of large DNA fragment. PMID:27364376

  10. Current Protocols in Mouse Biology Tissue-specific regulation of oncogene expression using Cre-inducible ROSA26 knock-in transgenic mice

    PubMed Central

    Carofino, Brandi L.; Justice, Monica J.

    2015-01-01

    Cre-inducible mouse models are often utilized for the spatial and temporal expression of oncogenes. With the wide number of Cre recombinase lines available, inducible transgenesis represents a tractable approach to achieve discrete oncogene expression. Here, we describe a protocol for targeting Cre-inducible genes using a loxP-STOP-loxP approach to the ubiquitously expressed ROSA26 locus. Gene targeting provides several advantages over standard transgenic techniques, including a known site of integration and previously characterized pattern of expression. Historically, an inherent instability of ROSA26 targeting vectors has hampered the efficiency of developing ROSA26 knock-in lines. In this protocol, we provide individual steps for utilizing Gateway recombination for cloning, and detailed instructions for screening targeted ES cell clones. By following this protocol, one can achieve germline transmission of a ROSA26 knock-in line within several months. PMID:26069083

  11. Strategic Planning for School Districts.

    ERIC Educational Resources Information Center

    Brown, Patricia; Marshall, Margaret

    Strategic planning is a process designed to move an educational organization through the steps of understanding changes in the external environment, assessing organizational strengths and weaknesses, developing a vision of a desired future and ways to achieve that mission, developing and implementing specific plans, and motivating that…

  12. Strategic Change in AAS Publishing

    NASA Astrophysics Data System (ADS)

    Steffen, Julie

    2015-08-01

    The American Astronomical Society has embarked on a process of strategic change in its publishing program. The process has incuded authors, AAS leaders, editors, publishing experts, librarians, and data scientists. This session will outline the still ongoing process and present some both upcoming and already available new AAS Publishing features and services to the global astronomy community.

  13. Entrepreneurial Spirit in Strategic Planning.

    ERIC Educational Resources Information Center

    Riggs, Donald E.

    1987-01-01

    Presents a model which merges the concepts of entrepreneurship with those of strategic planning to create a library management system. Each step of the process, including needs assessment and policy formation, strategy choice and implementation, and evaluation, is described in detail. (CLB)

  14. Strategic Audit and Marketing Plan

    ERIC Educational Resources Information Center

    Wright, Lianna S.

    2013-01-01

    The purpose of this audit was to revise the marketing plan for ADSum Professional Development School and give the owner a long-term vision of the school to operate competitively in the crowded field of for-profit schools. It is fairly simple to create a strategic plan but harder to implement and execute. Execution requires weeks and months of…

  15. Powerful Metrics: Strategic and Transformative

    ERIC Educational Resources Information Center

    Butterfield, Barbara

    2006-01-01

    To be a valuable partner at the strategic level, human resources can and should contribute to both institutional effectiveness measurement and workforce metrics. In this article, the author examines how to link HR initiatives with key institutional strategies, clarifies essential HR responsibilities for workforce results, explores return on human…

  16. Consultancy on Strategic Information Planning.

    ERIC Educational Resources Information Center

    Pejova, Zdravka, Ed.; Horton, Forest W., Ed.

    At the workshop, better management through strategic planning of information and consultancy was discussed as one way in which developing and Eastern European countries could tackle the complex information problems they are facing during the transition to a market economy. The sixteen papers in this volume are grouped into three basic categories:…

  17. Strategic petroleum reserve annual report

    SciTech Connect

    1996-02-15

    Section 165 of the Energy Policy and Conservation Act (Public Law 94- 163), as amended, requires the Secretary of Energy to submit annual reports to the President and the Congress on activities of the Strategic Petroleum Reserve (SPR). This report describes activities for the year ending December 31, 1995.

  18. Strategic development: a new focus.

    PubMed

    Lefko, J J

    1989-03-01

    Despite popular convention, a health-care organization's chief executive officer should be the organization's chief strategist. Planners and marketers must begin to function as strategic development officers responsible for sending the right information to the right decision makers so that they can make the right decisions as quickly as possible. PMID:10292364

  19. Strategic Planning of Technology Transfer.

    ERIC Educational Resources Information Center

    Groff, Warren H.

    Using the Ohio Technology Transfer Organization (OTTO) as its primary example, this paper offers a strategic planning perspective on technology transfer and human resources development. First, a brief overview is provided of the maturation of mission priorities and planning processes in higher education in the United States, followed by a…

  20. NASA Strategic Roadmap Summary Report

    NASA Technical Reports Server (NTRS)

    Wilson, Scott; Bauer, Frank; Stetson, Doug; Robey, Judee; Smith, Eric P.; Capps, Rich; Gould, Dana; Tanner, Mike; Guerra, Lisa; Johnston, Gordon

    2005-01-01

    In response to the Vision, NASA commissioned strategic and capability roadmap teams to develop the pathways for turning the Vision into a reality. The strategic roadmaps were derived from the Vision for Space Exploration and the Aldrich Commission Report dated June 2004. NASA identified 12 strategic areas for roadmapping. The Agency added a thirteenth area on nuclear systems because the topic affects the entire program portfolio. To ensure long-term public visibility and engagement, NASA established a committee for each of the 13 areas. These committees - made up of prominent members of the scientific and aerospace industry communities and senior government personnel - worked under the Federal Advisory Committee Act. A committee was formed for each of the following program areas: 1) Robotic and Human Lunar Exploration; 2) Robotic and Human Exploration of Mars; 3) Solar System Exploration; 4) Search for Earth-Like Planets; 5) Exploration Transportation System; 6) International Space Station; 7) Space Shuttle; 8) Universe Exploration; 9) Earth Science and Applications from Space; 10) Sun-Solar System Connection; 11) Aeronautical Technologies; 12) Education; 13) Nuclear Systems. This document contains roadmap summaries for 10 of these 13 program areas; The International Space Station, Space Shuttle, and Education are excluded. The completed roadmaps for the following committees: Robotic and Human Exploration of Mars; Solar System Exploration; Search for Earth-Like Planets; Universe Exploration; Earth Science and Applications from Space; Sun-Solar System Connection are collected in a separate Strategic Roadmaps volume. This document contains memebership rosters and charters for all 13 committees.

  1. Strategic Plan 2013 to 2016

    ERIC Educational Resources Information Center

    Lumina Foundation for Education, 2013

    2013-01-01

    In 2009, Lumina Foundation released its first strategic plan, based on the goal that 60% of Americans obtain a high-quality postsecondary degree or credential by 2025-- a goal now known as Goal 2025. Much has changed even in the short time since that plan was written, both in the external environment and in what has been learned from this work.…

  2. Strategic Planning for School Success.

    ERIC Educational Resources Information Center

    Herman, Jerry J.

    1993-01-01

    Strategic planners concerned with such matters as high-achieving students, high-performing teachers, broad-based community support, and a two-way involvement with the community must analyze the strengths, weaknesses, opportunities, and threats existing in the school's internal and external environment. A sample SWOT analysis is included. (MLH)

  3. Strategic Listening for School Leaders

    ERIC Educational Resources Information Center

    Tate, Jeannine S.; Dunklee, Dennis R.

    2005-01-01

    The ability to communicate effectively with multiple constituencies is recognized as an essential characteristic of effective leaders. Listening strategically is a way of showing parents, students, faculty, staff, and others that their ideas and beliefs are of value. The authors' practitioner-friendly book concentrates on the importance of…

  4. Strategic Planning: The Catastrophic Approach.

    ERIC Educational Resources Information Center

    Brody, Stanley J.

    1987-01-01

    Reviews and defines continuity of care, or long term care, as a potential economic catastrophe which requires strategic planning. Proposes a Medicare revision that provides integrated funding for acute, sub-acute, and long-term care as elements of continuity of care. (Author/ABB)

  5. Strategic School Planning in Jordan

    ERIC Educational Resources Information Center

    Al-Zboon, Mohammad Saleem; Hasan, Manal Subhi

    2012-01-01

    The study aimed to measuring the applying degree of the strategic school planning stages at the Governmental high schools from the educational supervisors and principals perspective in the directorates related to Amman city, the study society was formed of the educational supervisors and principals working at Educational directorates related to…

  6. How To Institutionalize Strategic Planning.

    ERIC Educational Resources Information Center

    Andrade, Sally J.

    1999-01-01

    The University of Texas at El Paso instituted a strategic planning framework with the intent of expanding and maintaining long-term participation by faculty, staff, and other constituents. The four-phase process included generating more open dialog about the future; responding to a powerful external incentive for self-assessment; preparing for…

  7. Strategic Approaches with Resistant Families.

    ERIC Educational Resources Information Center

    Breit, Miranda; And Others

    1983-01-01

    Describes the operation of a 10-session brief therapy unit for families who have failed in more traditional treatment modalities. Case material is presented to exemplify five different treatment strategies: symptom prescription, reframing, illusion of alternatives, role play, and strategic alliances. Advantages and limitations are discussed.…

  8. Strategic Planning in Distance Education

    ERIC Educational Resources Information Center

    Kilfoil, Wendy R.

    2003-01-01

    The purpose of strategic planning is to align the institution with the external environment. In fact, it is a complex solution to a complex problem. Environmental change is inevitable and it will always impact one's institution sooner or later. It is better to be proactive, thoughtful, and in charge of change, than to be always in a reactive mode…

  9. Strategic Planning for Technological Change

    ERIC Educational Resources Information Center

    Strong, Bart

    2007-01-01

    Strategic planning can become downright treacherous if one's institution depends heavily on technology, because technology changes course frequently. Technological change is somewhat predictable, and doing nothing is not an option. A number of complex factors hasten technological change. Limits on performance, breakthrough technology, market…

  10. Characteristics of Useful and Practical Organizational Strategic Plans

    ERIC Educational Resources Information Center

    Kaufman, Roger

    2014-01-01

    Most organizational strategic plans are not strategic but rather tactical or operational plans masquerading as "strategic." This article identifies the basic elements required in a useful and practical strategic plan and explains why they are important.

  11. 75 FR 22561 - Federal Advisory Committee; United States Strategic Command Strategic Advisory Group; Charter...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-29

    ... of the Secretary Federal Advisory Committee; United States Strategic Command Strategic Advisory Group... notice that it is renewing the charter for the United States Strategic Command Strategic Advisory Group... Management Officer for the Department of Defense, 703-601-6128. SUPPLEMENTARY INFORMATION: The Group is...

  12. Strategic Alliance Poker: Demonstrating the Importance of Complementary Resources and Trust in Strategic Alliance Management

    ERIC Educational Resources Information Center

    Reutzel, Christopher R.; Worthington, William J.; Collins, Jamie D.

    2012-01-01

    Strategic Alliance Poker (SAP) provides instructors with an opportunity to integrate the resource based view with their discussion of strategic alliances in undergraduate Strategic Management courses. Specifically, SAP provides Strategic Management instructors with an experiential exercise that can be used to illustrate the value creation…

  13. Metabolic Flux Analysis of Escherichia coli creB and arcA Mutants Reveals Shared Control of Carbon Catabolism under Microaerobic Growth Conditions▿

    PubMed Central

    Nikel, Pablo I.; Zhu, Jiangfeng; San, Ka-Yiu; Méndez, Beatriz S.; Bennett, George N.

    2009-01-01

    Escherichia coli has several elaborate sensing mechanisms for response to availability of oxygen and other electron acceptors, as well as the carbon source in the surrounding environment. Among them, the CreBC and ArcAB two-component signal transduction systems are responsible for regulation of carbon source utilization and redox control in response to oxygen availability, respectively. We assessed the role of CreBC and ArcAB in regulating the central carbon metabolism of E. coli under microaerobic conditions by means of 13C-labeling experiments in chemostat cultures of a wild-type strain, ΔcreB and ΔarcA single mutants, and a ΔcreB ΔarcA double mutant. Continuous cultures were conducted at D = 0.1 h−1 under carbon-limited conditions with restricted oxygen supply. Although all experimental strains metabolized glucose mainly through the Embden-Meyerhof-Parnas pathway, mutant strains had significantly lower fluxes in both the oxidative and the nonoxidative pentose phosphate pathways. Significant differences were also found at the pyruvate branching point. Both pyruvate-formate lyase and the pyruvate dehydrogenase complex contributed to acetyl-coenzyme A synthesis from pyruvate, and their activity seemed to be modulated by both ArcAB and CreBC. Strains carrying the creB deletion showed a higher biomass yield on glucose compared to the wild-type strain and its ΔarcA derivative, which also correlated with higher fluxes from building blocks to biomass. Glyoxylate shunt and lactate dehydrogenase were active mainly in the ΔarcA strain. Finally, it was observed that the tricarboxylic acid cycle reactions operated in a rather cyclic fashion under our experimental conditions, with reduced activity in the mutant strains. PMID:19561129

  14. 12 CFR 228.27 - Strategic plan.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 12 Banks and Banking 3 2013-01-01 2013-01-01 false Strategic plan. 228.27 Section 228.27 Banks and...) COMMUNITY REINVESTMENT (REGULATION BB) Standards for Assessing Performance § 228.27 Strategic plan. (a... assessment area(s) under a strategic plan if: (1) The bank has submitted the plan to the Board as...

  15. 12 CFR 228.27 - Strategic plan.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 12 Banks and Banking 3 2011-01-01 2011-01-01 false Strategic plan. 228.27 Section 228.27 Banks and... REINVESTMENT (REGULATION BB) Standards for Assessing Performance § 228.27 Strategic plan. (a) Alternative...(s) under a strategic plan if: (1) The bank has submitted the plan to the Board as provided for...

  16. 23 CFR 1335.6 - Strategic plan.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 23 Highways 1 2012-04-01 2012-04-01 false Strategic plan. 1335.6 Section 1335.6 Highways NATIONAL HIGHWAY TRAFFIC SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION STATE HIGHWAY SAFETY DATA IMPROVEMENTS § 1335.6 Strategic plan. A strategic plan shall— (a) Be a multi-year plan that identifies and...

  17. 12 CFR 563e.27 - Strategic plan.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 12 Banks and Banking 6 2014-01-01 2012-01-01 true Strategic plan. 563e.27 Section 563e.27 Banks... for Assessing Performance § 563e.27 Strategic plan. (a) Alternative election. The OTS will assess a... strategic plan if: (1) The savings association has submitted the plan to the OTS as provided for in...

  18. 12 CFR 563e.27 - Strategic plan.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 12 Banks and Banking 6 2012-01-01 2012-01-01 false Strategic plan. 563e.27 Section 563e.27 Banks... for Assessing Performance § 563e.27 Strategic plan. (a) Alternative election. The OTS will assess a... strategic plan if: (1) The savings association has submitted the plan to the OTS as provided for in...

  19. 13 CFR 313.6 - Strategic Plans.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 13 Business Credit and Assistance 1 2011-01-01 2011-01-01 false Strategic Plans. 313.6 Section 313... § 313.6 Strategic Plans. (a) General. An Impacted Community that intends to apply for a grant for implementation assistance under § 313.7 shall develop and submit a Strategic Plan to EDA for evaluation...

  20. 12 CFR 345.27 - Strategic plan.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 12 Banks and Banking 5 2012-01-01 2012-01-01 false Strategic plan. 345.27 Section 345.27 Banks and... REINVESTMENT Standards for Assessing Performance § 345.27 Strategic plan. (a) Alternative election. The FDIC... strategic plan if: (1) The bank has submitted the plan to the FDIC as provided for in this section; (2)...

  1. 12 CFR 228.27 - Strategic plan.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 12 Banks and Banking 3 2014-01-01 2014-01-01 false Strategic plan. 228.27 Section 228.27 Banks and...) COMMUNITY REINVESTMENT (REGULATION BB) Standards for Assessing Performance § 228.27 Strategic plan. (a... assessment area(s) under a strategic plan if: (1) The bank has submitted the plan to the Board as...

  2. 23 CFR 1335.6 - Strategic plan.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 23 Highways 1 2011-04-01 2011-04-01 false Strategic plan. 1335.6 Section 1335.6 Highways NATIONAL HIGHWAY TRAFFIC SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION STATE HIGHWAY SAFETY DATA IMPROVEMENTS § 1335.6 Strategic plan. A strategic plan shall— (a) Be a multi-year plan that identifies and...

  3. 23 CFR 1335.6 - Strategic plan.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 23 Highways 1 2010-04-01 2010-04-01 false Strategic plan. 1335.6 Section 1335.6 Highways NATIONAL HIGHWAY TRAFFIC SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION STATE HIGHWAY SAFETY DATA IMPROVEMENTS § 1335.6 Strategic plan. A strategic plan shall— (a) Be a multi-year plan that identifies and...

  4. Overview of Strategic Planning. Administrative Report.

    ERIC Educational Resources Information Center

    Raze, Nasus

    Strategic planning is a process by which the guiding members of an organization envision its future and develop tactics to achieve that future. This paper discusses the nature and purposes of strategic planning in educational organizations, and presents a 10-step generic model to illustrate the basic components of strategic planning. These 10…

  5. A Strategic Plan Is Just the Beginning

    ERIC Educational Resources Information Center

    Wilson, E. B.

    2009-01-01

    The process of strategic planning wears out institutions. Strategic planning, pursued with high purpose and energy, is enervating and exhausting. In this article, the author describes the value-added work of trustees in high-performing boards and discusses the role boards of trustees play as the process of continuous strategic planning unfolds.…

  6. Collaborative Strategic Planning: Myth or Reality?

    ERIC Educational Resources Information Center

    Mbugua, Flora; Rarieya, Jane F. A.

    2014-01-01

    The concept and practice of strategic planning, while entrenched in educational institutions in the West, is just catching on in Kenya. While literature emphasizes the importance of collaborative strategic planning, it does not indicate the challenges presented by collaboratively engaging in strategic planning. This article reports on findings of…

  7. Rethinking Strategy and Strategic Leadership in Schools.

    ERIC Educational Resources Information Center

    Davies, Brent

    2003-01-01

    Reviews nature of strategy and strategic leadership in schools. Considers how leaders can map and reconceptualize the nature of strategy and develop strategic capabilities for longer-term sustainability. Questions hierarchical models of leadership. Highlights three characteristics of strategically oriented schools; suggests ways to improve art of…

  8. Data and information quality strategic plan.

    PubMed

    Bethell, C

    The Environmental Protection Agency's Strategic Plan was developed in response to internal and external concerns about the integrity, consistency, and accuracy of EPA's environmental data. This document explains why a Strategic Plan is needed and the methodology used in its development, cites Agency models of excellence, and presents the six recommendations of EPA's Data and Information Quality Strategic Plan. PMID:12512550

  9. Strategic Activism, Educational Leadership and Social Justice

    ERIC Educational Resources Information Center

    Ryan, James

    2016-01-01

    This article describes the strategic activism of educational leaders who promote social justice. Given the risks, educational leaders need to be strategic about the ways in which they pursue their activism. Citing current research, this article explores the ways in which leaders strategically pursue their social justice agendas within their own…

  10. Putative methyltransferase LaeA and transcription factor CreA are necessary for proper asexual development and controlling secondary metabolic gene cluster expression.

    PubMed

    Zhang, Xiujun; Zhu, Yingying; Bao, Longfei; Gao, Liwei; Yao, Guangshan; Li, Yanan; Yang, Zhifeng; Li, Zhonghai; Zhong, Yaohua; Li, Fuli; Yin, Heng; Qu, Yinbo; Qin, Yuqi

    2016-09-01

    The morphological development of fungi is a complex process and is often coupled with secondary metabolite production. In this study, we assessed the function of putative methyltransferase LaeA and transcription factor CreA in controlling asexual development and secondary metabolic gene cluster expression in Penicillium oxalicum. The deletion of laeA (ΔlaeA) impaired the conidiation in P. oxalicum, with a downregulated expression of brlA. Overexpression of P. oxalicum brlA in ΔlaeA could upregulate brlA and abaA remarkably, but could not rescue the conidiation defect; therefore, brlA and abaA expression were necessary but not sufficient for conidiation. Deletion of creA in ΔlaeA background (ΔlaeAΔcreA) blocked conidiation with a white fluffy phenotype. Nutrient-rich medium could not rescue developmental defects in ΔlaeAΔcreA mutant but could rescue defects in ΔlaeA. Expression of 10 genes, namely, albA/wA, abrB/yA, arpA, aygA, arpA-like, arpB, arpB-like, rodA, rodA-like, and rodB, for pigmentation and spore wall protein genes was silenced in ΔlaeAΔcreA, whereas only six of them were downregulated in ΔlaeA. Among the 28 secondary metabolism gene clusters in P. oxalicum, four secondary metabolism gene clusters were silenced in ΔlaeA and two were also silenced in ΔbrlA mutant. A total of 10 physically linked and coregulated genes were distributed over five chromosomes in ΔlaeA. Six of these genes were located in subtelomeric regions, thus demonstrating a positional bias for LaeA-regulated clusters toward subtelomeric regions. All of silenced clusters located in subtelomeric regions were derepressed in ΔlaeAΔcreA, hence showing that lack of CreA could remediate the repression of gene clusters in ΔlaeA background. Results show that both putative methyltransferase LaeA and transcription factor CreA are necessary for proper asexual development and controlling secondary metabolic gene cluster expression. PMID:27387217

  11. Vesicular demyelination induced by raised intracellular calcium.

    PubMed

    Smith, K J; Hall, S M; Schauf, C L

    1985-11-01

    myelin vesiculation. We conclude that vesicular demyelination can be initiated in vital Schwann cells by a raised intracellular Ca2+ concentration. Such demyelination does not necessarily lead to Schwann cell death. The possible relevance of the findings to vesicular demyelinating neuropathies is discussed, and a hypothesis regarding the mechanism of demyelination is advanced. PMID:3003255

  12. Proglucagon Promoter Cre-Mediated AMPK Deletion in Mice Increases Circulating GLP-1 Levels and Oral Glucose Tolerance

    PubMed Central

    Sayers, Sophie R.; Reimann, Frank; Gribble, Fiona M.; Parker, Helen; Zac-Varghese, Sagen; Bloom, Stephen R.; Foretz, Marc; Viollet, Benoit; Rutter, Guy A.

    2016-01-01

    Background Enteroendocrine L-cells synthesise and release the gut hormone glucagon-like peptide-1 (GLP-1) in response to food transit. Deletion of the tumour suppressor kinase LKB1 from proglucagon-expressing cells leads to the generation of intestinal polyps but no change in circulating GLP-1 levels. Here, we explore the role of the downstream kinase AMP-activated protein kinase (AMPK) in these cells. Method Loss of AMPK from proglucagon-expressing cells was achieved using a preproglucagon promoter-driven Cre (iGluCre) to catalyse recombination of floxed alleles of AMPKα1 and α2. Oral and intraperitoneal glucose tolerance were measured using standard protocols. L-cell mass was measured by immunocytochemistry. Hormone and peptide levels were measured by electrochemical-based luminescence detection or radioimmunoassay. Results Recombination with iGluCre led to efficient deletion of AMPK from intestinal L- and pancreatic alpha-cells. In contrast to mice rendered null for LKB1 using the same strategy, mice deleted for AMPK displayed an increase (WT: 0.05 ± 0.01, KO: 0.09±0.02%, p<0.01) in L-cell mass and elevated plasma fasting (WT: 5.62 ± 0.800 pg/ml, KO: 14.5 ± 1.870, p<0.01) and fed (WT: 15.7 ± 1.48pg/ml, KO: 22.0 ± 6.62, p<0.01) GLP-1 levels. Oral, but not intraperitoneal, glucose tolerance was significantly improved by AMPK deletion, whilst insulin and glucagon levels were unchanged despite an increase in alpha to beta cell ratio (WT: 0.23 ± 0.02, KO: 0.33 ± 0.03, p<0.01). Conclusion AMPK restricts L-cell growth and GLP-1 secretion to suppress glucose tolerance. Targeted inhibition of AMPK in L-cells may thus provide a new therapeutic strategy in some forms of type 2 diabetes. PMID:27010458

  13. Developing strategic thinking in senior management.

    PubMed

    Zabriskie, N B; Huellmantel, A B

    1991-12-01

    Chief Executive Officers have recently stated that their greatest staffing challenge for the 1990s is the development of strategic leadership in their senior management. In order to do this, it is necessary to identify the substance of strategic thinking, and the capabilities that must be mastered. Writers on strategy have identified six major elements of strategic thinking and these have been organized to reveal the tasks, questions, decisions, and skills that senior executives must acquire in order to lead their organizations strategically. Finally, the article identifies training programme elements which are used by Directors of Manpower Development to develop strategic leadership ability. PMID:10118696

  14. Analysis of lung tumor initiation and progression in transgenic mice for Cre-inducible overexpression of Cul4A gene

    DOE PAGESBeta

    Wang, Yang; Xu, Zhidong; Mao, Jian -Hua; Hung, Ming -Szu; Hsieh, David; Au, Alfred; Jablons, David M.; You, Liang

    2015-06-08

    Background: Lung cancer is the leading cause of morbidity and death worldwide. Although the available lung cancer animal models have been informative and further propel our understanding of human lung cancer, they still do not fully recapitulate the complexities of human lung cancer. The pathogenesis of lung cancer remains highly elusive because of its aggressive biologic nature and considerable heterogeneity, compared to other cancers. The association of Cul4A amplification with aggressive tumor growth and poor prognosis has been suggested. Our previous study suggested that Cul4A is oncogenic in vitro, but its oncogenic role in vivo has not been studied. Methods:more » Viral delivery approaches have been used extensively to model cancer in mouse models. In our experiments, we used Cre-recombinase induced overexpression of the Cul4A gene in transgenic mice to study the role of Cul4A on lung tumor initiation and progression and have developed a new model of lung tumor development in mice harboring a conditionally expressed allele of Cul4A. Results: Here we show that the use of a recombinant adenovirus expressing Cre-recombinase (“AdenoCre”) to induce Cul4A overexpression in the lungs of mice allows controls of the timing and multiplicity of tumor initiation. Following our mouse models, we are able to study the potential role of Cul4A in the development and progression in pulmonary adenocarcinoma as well. Conclusion: Our findings indicate that Cul4A is oncogenic in vivo, and this mouse model is a tool in understanding the mechanisms of Cul4A in human cancers and for testing experimental therapies targeting Cul4A.« less

  15. Analysis of lung tumor initiation and progression in transgenic mice for Cre-inducible overexpression of Cul4A gene

    SciTech Connect

    Wang, Yang; Xu, Zhidong; Mao, Jian -Hua; Hung, Ming -Szu; Hsieh, David; Au, Alfred; Jablons, David M.; You, Liang

    2015-06-08

    Background: Lung cancer is the leading cause of morbidity and death worldwide. Although the available lung cancer animal models have been informative and further propel our understanding of human lung cancer, they still do not fully recapitulate the complexities of human lung cancer. The pathogenesis of lung cancer remains highly elusive because of its aggressive biologic nature and considerable heterogeneity, compared to other cancers. The association of Cul4A amplification with aggressive tumor growth and poor prognosis has been suggested. Our previous study suggested that Cul4A is oncogenic in vitro, but its oncogenic role in vivo has not been studied. Methods: Viral delivery approaches have been used extensively to model cancer in mouse models. In our experiments, we used Cre-recombinase induced overexpression of the Cul4A gene in transgenic mice to study the role of Cul4A on lung tumor initiation and progression and have developed a new model of lung tumor development in mice harboring a conditionally expressed allele of Cul4A. Results: Here we show that the use of a recombinant adenovirus expressing Cre-recombinase (“AdenoCre”) to induce Cul4A overexpression in the lungs of mice allows controls of the timing and multiplicity of tumor initiation. Following our mouse models, we are able to study the potential role of Cul4A in the development and progression in pulmonary adenocarcinoma as well. Conclusion: Our findings indicate that Cul4A is oncogenic in vivo, and this mouse model is a tool in understanding the mechanisms of Cul4A in human cancers and for testing experimental therapies targeting Cul4A.

  16. A 3D Searchable Database of Transgenic Zebrafish Gal4 and Cre Lines for Functional Neuroanatomy Studies.

    PubMed

    Marquart, Gregory D; Tabor, Kathryn M; Brown, Mary; Strykowski, Jennifer L; Varshney, Gaurav K; LaFave, Matthew C; Mueller, Thomas; Burgess, Shawn M; Higashijima, Shin-Ichi; Burgess, Harold A

    2015-01-01

    Transgenic methods enable the selective manipulation of neurons for functional mapping of neuronal circuits. Using confocal microscopy, we have imaged the cellular-level expression of 109 transgenic lines in live 6 day post fertilization larvae, including 80 Gal4 enhancer trap lines, 9 Cre enhancer trap lines and 20 transgenic lines that express fluorescent proteins in defined gene-specific patterns. Image stacks were acquired at single micron resolution, together with a broadly expressed neural marker, which we used to align enhancer trap reporter patterns into a common 3-dimensional reference space. To facilitate use of this resource, we have written software that enables searching for transgenic lines that label cells within a selectable 3-dimensional region of interest (ROI) or neuroanatomical area. This software also enables the intersectional expression of transgenes to be predicted, a feature which we validated by detecting cells with co-expression of Cre and Gal4. Many of the imaged enhancer trap lines show intrinsic brain-specific expression. However, to increase the utility of lines that also drive expression in non-neuronal tissue we have designed a novel UAS reporter, that suppresses expression in heart, muscle, and skin through the incorporation of microRNA binding sites in a synthetic 3' untranslated region. Finally, we mapped the site of transgene integration, thus providing molecular identification of the expression pattern for most lines. Cumulatively, this library of enhancer trap lines provides genetic access to 70% of the larval brain and is therefore a powerful and broadly accessible tool for the dissection of neural circuits in larval zebrafish. PMID:26635538

  17. A 3D Searchable Database of Transgenic Zebrafish Gal4 and Cre Lines for Functional Neuroanatomy Studies

    PubMed Central

    Marquart, Gregory D.; Tabor, Kathryn M.; Brown, Mary; Strykowski, Jennifer L.; Varshney, Gaurav K.; LaFave, Matthew C.; Mueller, Thomas; Burgess, Shawn M.; Higashijima, Shin-ichi; Burgess, Harold A.

    2015-01-01

    Transgenic methods enable the selective manipulation of neurons for functional mapping of neuronal circuits. Using confocal microscopy, we have imaged the cellular-level expression of 109 transgenic lines in live 6 day post fertilization larvae, including 80 Gal4 enhancer trap lines, 9 Cre enhancer trap lines and 20 transgenic lines that express fluorescent proteins in defined gene-specific patterns. Image stacks were acquired at single micron resolution, together with a broadly expressed neural marker, which we used to align enhancer trap reporter patterns into a common 3-dimensional reference space. To facilitate use of this resource, we have written software that enables searching for transgenic lines that label cells within a selectable 3-dimensional region of interest (ROI) or neuroanatomical area. This software also enables the intersectional expression of transgenes to be predicted, a feature which we validated by detecting cells with co-expression of Cre and Gal4. Many of the imaged enhancer trap lines show intrinsic brain-specific expression. However, to increase the utility of lines that also drive expression in non-neuronal tissue we have designed a novel UAS reporter, that suppresses expression in heart, muscle, and skin through the incorporation of microRNA binding sites in a synthetic 3′ untranslated region. Finally, we mapped the site of transgene integration, thus providing molecular identification of the expression pattern for most lines. Cumulatively, this library of enhancer trap lines provides genetic access to 70% of the larval brain and is therefore a powerful and broadly accessible tool for the dissection of neural circuits in larval zebrafish. PMID:26635538

  18. Strategic leadership: the essential skills.

    PubMed

    Schoemaker, Paul J H; Krupp, Steve; Howland, Samantha

    2013-01-01

    The more uncertain your environment, the greater the opportunity--if you have the leadership skills to capitalize on it. Research at the Wharton school and at the authors' consulting firm, involving more than 20,000 executives to date, has identified six skills that, when mastered and used in concert, allow leaders to think strategically and navigate the unknown effectively. They are the abilities to anticipate, challenge, interpret, decide, align, and learn. This article describes the six skills in detail and includes a self-assessment that will enable you to identify the ones that most need your attention. The authors have found that strength in one skill cannot easily compensate for a deficit in another. An adaptive strategic leader has learned to apply all six at once. PMID:23390746

  19. Strategic planning for every manager.

    PubMed

    Reeves, P N

    1990-01-01

    All managers plan, but many managers of small organizations limit their planning activities to developing and guiding discrete projects. This decision stems from two erroneous assumptions: strategic planning is useful only in large organizations, and it is too complex for a small organization. As a result, these managers deprive their organizations of an essential tool for long-term survival and prosperity. Strategic planning helps any organization effectively use its scarce resources by relating every decision to the organization's mission. The process for doing this consists of a sequence of decisions that applies in all cases. However, the effort involved in reaching each of these decisions varies with the size and complexity of the organization. Consequently, leaders of small units need not forego using this essential managerial tool. In this article, each decision is described and illustrated with an example from a typical small enterprise. PMID:10106892

  20. Building a strategic security organisation.

    PubMed

    Howard, Mike

    2016-01-01

    In everyone's day-to-day jobs there is constant need to deal with current and newly detected matters. This is now a world of immediacy, driven by the cadence of the business and its needs. These concerns should not be ignored, as failing to deal with these issues would not bode well for the future. It is essential that the gears are kept spinning. The challenge for any security organisation is to identify its short-term tactical requirements, while developing longer-term strategic needs. Once done, the differences can be accounted for and strides can be made toward a desired future state. This paper highlights several steps that the author and his team have taken in their own journey. There is no magic answer, each organisation will have its own unique challenges. Nevertheless, some of the approaches to building a strategic security organisation described in this paper are applicable to all organisations, irrespective of their size. PMID:27318284

  1. Strategic alliances and market risk.

    PubMed

    Havenaar, Matthias; Hiscocks, Peter

    2012-08-01

    Strategic alliances in product development and marketing are crucial to the biotechnology industry. Many alliances, however, are terminated before the drug reaches the market. In this article we make the case that strategic alliances can fail because of how they are negotiated. Alliance contracts are often inflexible and do not allow for changes in market conditions. We propose a model for contract valuation that can assist biotech and/or pharma deal makers in negotiating alliances that have a higher chance of survival in uncertain market conditions. The model makes use of variable royalties and milestone payments. Because licensing is key to the biotech and/or pharma business model this article will be of interest not only to professionals in licensing, but to all professionals active in the industry. PMID:22484547

  2. Strategic Defense Initiative - an overview

    SciTech Connect

    Abrahamson, J.A.

    1985-01-01

    A general framework for developing the concept and implementing options for strategic ballistic missile defense systems is emerging. The objective of the Strategic Defense Initiative (SDI) program is to conduct research on those technologies for defensive system which could intercept ballistic missiles after they have been launched and prevent them from hitting their targets. Its goal is to acquire the technical knowledge as a basis for a later decision on deployment, not a program for deployment or star wars. Abrahamson reviews the basic technological questions facing researchers, outlines the multi-layered defense possibilities and summarizes the status of technology to date. He argues for a continuity of resources to conduct the program. 4 figures.

  3. Eye Movements in Strategic Choice

    PubMed Central

    Gächter, Simon; Noguchi, Takao; Mullett, Timothy L.

    2015-01-01

    Abstract In risky and other multiattribute choices, the process of choosing is well described by random walk or drift diffusion models in which evidence is accumulated over time to threshold. In strategic choices, level‐k and cognitive hierarchy models have been offered as accounts of the choice process, in which people simulate the choice processes of their opponents or partners. We recorded the eye movements in 2 × 2 symmetric games including dominance‐solvable games like prisoner's dilemma and asymmetric coordination games like stag hunt and hawk–dove. The evidence was most consistent with the accumulation of payoff differences over time: we found longer duration choices with more fixations when payoffs differences were more finely balanced, an emerging bias to gaze more at the payoffs for the action ultimately chosen, and that a simple count of transitions between payoffs—whether or not the comparison is strategically informative—was strongly associated with the final choice. The accumulator models do account for these strategic choice process measures, but the level‐k and cognitive hierarchy models do not. © 2015 The Authors. Journal of Behavioral Decision Making published by John Wiley & Sons Ltd.

  4. Strategic petroleum reserve. Quarterly report

    SciTech Connect

    1995-08-15

    The Strategic Petroleum Reserve reduces the Nation`s vulnerability to oil supply disruptions. Its existence provides a formidable deterrent to the use of oil as a political instrument and an effective response mechanism should a disruption occur. The Strategic Petroleum Reserve was created pursuant to the Energy Policy and Conservation Act of December 22, 1975 (Public Law 94-163). Its purposes are to reduce the impact of disruptions in supplies of petroleum products and to carry out obligations of the United States under the Agreement on an International Energy Program. Section 165(a) of the Act requires the submission of Annual Reports and Section 165(b)(1) requires the submission of Quarterly Reports. This Quarterly Report highlights activities undertaken during the second quarter of calendar year 1995, including: inventory of petroleum products stored in the Reserve; current and projected storage capacity, analysis of existing or anticipated problems with the acquisition and storage of petroleum products, and future expansion of storage capacity; funds obligated by the Secretary from the SPR Petroleum Account and the Strategic Petroleum Reserve Account during the prior calendar quarter and in total; and major environmental actions completed, in progress, or anticipated.

  5. Strategic petroleum reserve. Quarterly report

    SciTech Connect

    Not Available

    1994-05-15

    The Strategic Petroleum Reserve serves as one of our most important investments in reducing the Nation`s vulnerability to oil supply disruptions. Its existence provides an effective response mechanism should a disruption occur and a formidable deterrent to the use of oil as a political instrument. The Strategic Petroleum Reserve was created pursuant to the Energy Policy and Conservation Act of December 22, 1975, (Public Law 94-163) as amended, to reduce the impact of disruptions in supplies of petroleum products and to carry out obligations of the United States under the Agreement on an International Energy Program. Section 165(a) of the Act requires the submission of Annual Reports and Section 165(b)(1) requires the submission of Quarterly Reports. This Quarterly Report highlights activities undertaken during the first quarter of calendar year 1994, including: (1) inventory of petroleum products stored in the Reserve, under contract and in transit at the end of the calendar quarter; (2) fill rate for the current quarter and projected fill rate for the next calendar quarter; (3) average price of the petroleum products acquired during the calendar quarter; (4) current and projected storage capacity; (5) analysis of existing or anticipated problems with the acquisition and storage of petroleum products, and future expansion of storage capacity; (6) funds obligated by the Secretary from the SPR Petroleum Account and the Strategic Petroleum Reserve Account during the prior calendar quarter and in total; and (7) major environmental actions completed, in progress, or anticipated.

  6. Adenovirus-mediated Cre deletion of floxed sequences in primary mouse cells is an efficient alternative for studies of gene deletion

    PubMed Central

    Prost, Sandrine; Sheahan, Sharon; Rannie, Dominic; Harrison, David J.

    2001-01-01

    This study evaluates the utility of Cre-expressing adenovirus for deletion of floxed genes in primary cells using primary murine hepatocytes. Adenovirus infection was very efficient, even at very low MOI (>95% infection at a MOI of 6) and did not reduce viability. High level LacZ expression was cytotoxic to hepatocytes but Cre expression had no effect on viability. Cre-mediated recombination was completed within a timespan that permits experimentation during primary culture (>95% recombination after 24 h), independently of the number of floxed alleles per cell. Recombination did not induce p53 or produce cytological nuclear abnormalities (even in polyploid cells). Contrary to expectation, deletion of DNA ligase 1 did not alter cell cycle progression, although Cre expression hastens entry to S phase from G1, independently of the presence of floxed sequences. We conclude that adenovirus-mediated deletion of floxed alleles in primary cells is a straightforward and highly efficient tool for conducting preliminary studies of conditional gene targeting. Primary cells have advantages of differentiation, relative purity and ease of experimentation within controlled conditions, while avoiding confounding problems encountered in vivo (i.e. target cell specificity, kinetics and level of recombination, and elicitation of inflammatory and immune responses). This system could help identify important phenotypic effects and design and interpret in vivo studies. PMID:11504888

  7. Nucleo-cytoplasmic shuttling dynamics of the transcriptional regulators XYR1 and CRE1 under conditions of cellulase and xylanase gene expression in Trichoderma reesei

    PubMed Central

    Lichius, Alexander; Seidl-Seiboth, Verena; Seiboth, Bernhard; Kubicek, Christian P

    2014-01-01

    Trichoderma reesei is a model for investigating the regulation of (hemi-)cellulase gene expression. Cellulases are formed adaptively, and the transcriptional activator XYR1 and the carbon catabolite repressor CRE1 are main regulators of their expression. We quantified the nucleo-cytoplasmic shuttling dynamics of GFP-fusion proteins of both transcription factors under cellulase and xylanase inducing conditions, and correlated their nuclear presence/absence with transcriptional changes. We also compared their subcellular localization in conidial germlings and mature hyphae. We show that cellulase gene expression requires de novo biosynthesis of XYR1 and its simultaneous nuclear import, whereas carbon catabolite repression is regulated through preformed CRE1 imported from the cytoplasmic pool. Termination of induction immediately stopped cellulase gene transcription and was accompanied by rapid nuclear degradation of XYR1. In contrast, nuclear CRE1 rapidly decreased upon glucose depletion, and became recycled into the cytoplasm. In mature hyphae, nuclei containing activated XYR1 were concentrated in the colony center, indicating that this is the main region of XYR1 synthesis and cellulase transcription. CRE1 was found to be evenly distributed throughout the entire mycelium. Taken together, our data revealed novel aspects of the dynamic shuttling and spatial bias of the major regulator of (hemi-)cellulase gene expression, XYR1, in T. reesei. PMID:25302561

  8. Robust inducible Cre recombinase activity in the human malaria parasite Plasmodium falciparum enables efficient gene deletion within a single asexual erythrocytic growth cycle.

    PubMed

    Collins, Christine R; Das, Sujaan; Wong, Eleanor H; Andenmatten, Nicole; Stallmach, Robert; Hackett, Fiona; Herman, Jean-Paul; Müller, Sylke; Meissner, Markus; Blackman, Michael J

    2013-05-01

    Asexual blood stages of the malaria parasite, which cause all the pathology associated with malaria, can readily be genetically modified by homologous recombination, enabling the functional study of parasite genes that are not essential in this part of the life cycle. However, no widely applicable method for conditional mutagenesis of essential asexual blood-stage malarial genes is available, hindering their functional analysis. We report the application of the DiCre conditional recombinase system to Plasmodium falciparum, the causative agent of the most dangerous form of malaria. We show that DiCre can be used to obtain rapid, highly regulated site-specific recombination in P. falciparum, capable of excising loxP-flanked sequences from a genomic locus with close to 100% efficiency within the time-span of a single erythrocytic growth cycle. DiCre-mediated deletion of the SERA5 3' UTR failed to reduce expression of the gene due to the existence of alternative cryptic polyadenylation sites within the modified locus. However, we successfully used the system to recycle the most widely used drug resistance marker for P. falciparum, human dihydrofolate reductase, in the process producing constitutively DiCre-expressing P. falciparum clones that have broad utility for the functional analysis of essential asexual blood-stage parasite genes. PMID:23489321

  9. Development of a conditional liver tumor model by mifepristone-inducible Cre recombination to control oncogenic krasV12 expression in transgenic zebrafish

    PubMed Central

    Nguyen, Anh Tuan; Koh, Vivien; Spitsbergen, Jan M.; Gong, Zhiyuan

    2016-01-01

    Here we report a new transgenic expression system by combination of liver-specific expression, mifepristone induction and Cre-loxP recombination to conditionally control the expression of oncogenic krasV12. This transgenic system allowed expression of krasV12 specifically in the liver by a brief exposure of mifepristone to induce permanent genomic recombination mediated by the Cre-loxP system. We found that liver tumors were generally induced from multiple foci due to incomplete Cre-loxP recombination, thus mimicking naturally occurring human tumors resulting from one or a few mutated cells and clonal proliferation to form nodules. Similar to our earlier studies by both constitutive and inducible expression of the krasV12 oncogene, hepatocellular carcinoma (HCC) is the main type of liver tumor induced by krasV12 expression. Moreover, mixed tumors with hepatocellular adenoma and hepatoblastoma (HB) were also frequently observed. Molecular analyses also indicated similar increase of phosphorylated ERK1/2 in all types of liver tumors, but nuclear localization of β–catenin, a sign of malignant transformation, was found only in HCC and HB. Taken together, our new transgenic system reported in this study allows transgenic krasV12 expression specifically in the zebrafish liver only by a brief exposure of mifepristone to induce permanent genomic recombination mediated by the Cre-loxP system. PMID:26790949

  10. Conditional gene deletion with DiCre demonstrates an essential role for CRK3 in L eishmania mexicana cell cycle regulation

    PubMed Central

    Duncan, Samuel M.; Myburgh, Elmarie; Philipon, Cintia; Brown, Elaine; Meissner, Markus; Brewer, James

    2016-01-01

    Summary Leishmania mexicana has a large family of cyclin‐dependent kinases (CDKs) that reflect the complex interplay between cell cycle and life cycle progression. Evidence from previous studies indicated that Cdc2‐related kinase 3 (CRK3) in complex with the cyclin CYC6 is a functional homologue of the major cell cycle regulator CDK1, yet definitive genetic evidence for an essential role in parasite proliferation is lacking. To address this, we have implemented an inducible gene deletion system based on a dimerised Cre recombinase (diCre) to target CRK3 and elucidate its role in the cell cycle of L. mexicana. Induction of diCre activity in promastigotes with rapamycin resulted in efficient deletion of floxed CRK3, resulting in G2/M growth arrest. Co‐expression of a CRK3 transgene during rapamycin‐induced deletion of CRK3 resulted in complementation of growth, whereas expression of an active site CRK3 T178E mutant did not, showing that protein kinase activity is crucial for CRK3 function. Inducible deletion of CRK3 in stationary phase promastigotes resulted in attenuated growth in mice, thereby confirming CRK3 as a useful therapeutic target and diCre as a valuable new tool for analyzing essential genes in Leishmania. PMID:26991545

  11. aP2-Cre-mediated inactivation of acetyl-CoA carboxylase 1 causes growth retardation and reduced lipid accumulation in adipose tissues

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Adipose tissue is one of the major sites for fatty acid synthesis and lipid storage. We generated adipose (fat)-specific ACC1 knockout (FACC1KO) mice using the aP2-Cre/loxP system. FACC1KO mice showed prenatal growth retardation; after weaning, however, their weight gain was comparable to that of wi...

  12. Heat shock induced excision of selectable marker genes in transgenic banana by the Cre-lox site-specific recombination system.

    PubMed

    Chong-Pérez, Borys; Kosky, Rafael G; Reyes, Maritza; Rojas, Luis; Ocaña, Bárbara; Tejeda, Marisol; Pérez, Blanca; Angenon, Geert

    2012-06-30

    Selectable marker genes are indispensable for efficient production of transgenic events, but are no longer needed after the selection process and may cause public concern and technological problems. Although several gene excision systems exist, few have been optimized for vegetatively propagated crops. Using a Cre-loxP auto-excision strategy, we obtained transgenic banana plants cv. Grande Naine (Musa AAA) devoid of the marker gene used for selection. We used T-DNA vectors with the cre recombinase gene under control of a heat shock promoter and selectable marker gene cassettes placed between two loxP sites in direct orientation, and a gene of interest inserted outside of the loxP sites. Heat shock promoters pGmHSP17.6-L and pHSP18.2, from soybean and Arabidopsis respectively, were tested. A transient heat shock treatment of primary transgenic embryos was sufficient for inducing cre and excising cre and the marker genes. Excision efficiency, as determined by PCR and Southern hybridization was 59.7 and 40.0% for the GmHSP17.6-L and HSP18.2 promoters, respectively. Spontaneous excision was not observed in 50 plants derived from untreated transgenic embryos. To our knowledge this is the first report describing an efficient marker gene removal system for banana. The method described is simple and might be generally applicable for the production of marker-free transgenic plants of many crop species. PMID:21839123

  13. Generation of Pet1210-Cre Transgenic Mouse Line Reveals Non-Serotonergic Expression Domains of Pet1 Both in CNS and Periphery

    PubMed Central

    Pelosi, Barbara; Migliarini, Sara; Pacini, Giulia; Pratelli, Marta; Pasqualetti, Massimo

    2014-01-01

    Neurons producing serotonin (5-hydroxytryptamine, 5-HT) constitute one of the most widely distributed neuronal networks in the mammalian central nervous system (CNS) and exhibit a profuse innervation throughout the CNS already at early stages of development. Serotonergic neuron specification is controlled by a combination of secreted molecules and transcription factors such as Shh, Fgf4/8, Nkx2.2, Lmx1b and Pet1. In the mouse, Pet1 mRNA expression appears between 10 and 11 days post coitum (dpc) in serotonergic post-mitotic precursors and persists in serotonergic neurons up to adulthood, where it promotes the expression of genes defining the mature serotonergic phenotype such as tryptophan hydroxylase 2 (Tph2) and serotonin transporter (SERT). Hence, the generation of genetic tools based on Pet1 specific expression represents a valuable approach to study the development and function of the serotonergic system. Here, we report the generation of a Pet1210-Cre transgenic mouse line in which the Cre recombinase is expressed under the control of a 210 kb fragment from the Pet1 genetic locus to ensure a reliable and faithful control of somatic recombination in Pet1 cell lineage. Besides Cre-mediated recombination accurately occurred in the serotonergic system as expected and according to previous studies, Pet1210-Cre transgenic mouse line allowed us to identify novel, so far uncharacterized, Pet1 expression domains. Indeed, we showed that in the raphe Pet1 is expressed also in a non-serotonergic neuronal population intermingled with Tph2-expressing cells and mostly localized in the B8 and B9 nuclei. Moreover, we detected Cre-mediated recombination also in the developing pancreas and in the ureteric bud derivatives of the kidney, where it reflected a specific Pet1 expression. Thus, Pet1210-Cre transgenic mouse line faithfully drives Cre-mediated recombination in all Pet1 expression domains representing a valuable tool to genetically manipulate serotonergic and non

  14. Stress-Induced Anxiety- and Depressive-Like Phenotype Associated with Transient Reduction in Neurogenesis in Adult Nestin-CreERT2/Diphtheria Toxin Fragment A Transgenic Mice

    PubMed Central

    Yun, Sanghee; Donovan, Michael H.; Ross, Michele N.; Richardson, Devon R.; Reister, Robin; Farnbauch, Laure A.; Fischer, Stephanie J.; Riethmacher, Dieter; Gershenfeld, Howard K.; Lagace, Diane C.; Eisch, Amelia J.

    2016-01-01

    Depression and anxiety involve hippocampal dysfunction, but the specific relationship between these mood disorders and adult hippocampal dentate gyrus neurogenesis remains unclear. In both humans with MDD and rodent models of depression, administration of antidepressants increases DG progenitor and granule cell number, yet rodents with induced ablation of DG neurogenesis typically do not demonstrate depressive- or anxiety-like behaviors. The conflicting data may be explained by the varied duration and degree to which adult neurogenesis is reduced in different rodent neurogenesis ablation models. In order to test this hypothesis we examined how a transient–rather than permanent–inducible reduction in neurogenesis would alter depressive- and anxiety-like behaviors. Transgenic Nestin-CreERT2/floxed diphtheria toxin fragment A (DTA) mice (Cre+DTA+) and littermates (Cre+DTA-; control) were given tamoxifen (TAM) to induce recombination and decrease nestin-expressing stem cells and their progeny. The decreased neurogenesis was transient: 12 days post-TAM Cre+DTA+ mice had fewer DG proliferating Ki67+ cells and fewer DCX+ neuroblasts/immature neurons relative to control, but 30 days post-TAM Cre+DTA+ mice had the same DCX+ cell number as control. This ability of DG neurogenesis to recover after partial ablation also correlated with changes in behavior. Relative to control, Cre+DTA+ mice tested between 12–30 days post-TAM displayed indices of a stress-induced anxiety phenotype–longer latency to consume highly palatable food in the unfamiliar cage in the novelty-induced hypophagia test, and a depression phenotype–longer time of immobility in the tail suspension test, but Cre+DTA+ mice tested after 30 days post-TAM did not. These findings suggest a functional association between adult neurogenesis and stress induced anxiety- and depressive-like behaviors, where induced reduction in DCX+ cells at the time of behavioral testing is coupled with stress-induced anxiety

  15. Endogenous n-3 Polyunsaturated Fatty Acids Delay Progression of Pancreatic Ductal Adenocarcinoma in Fat-1-p48Cre/+-LSL-KrasG12D/+ Mice12

    PubMed Central

    Mohammed, Altaf; Janakiram, Naveena B; Brewer, Misty; Duff, Ashley; Lightfoot, Stan; Brush, Richard S; Anderson, Robert E; Rao, Chinthalapally V

    2012-01-01

    Preclinical studies suggest that diets rich in omega-3 polyunsaturated fatty acids (n-3 PUFAs) may be beneficial for prevention of pancreatic cancer. Nutritional intervention studies are often complex, and there is no clear evidence, without potential confounding factors, on whether conversion of n-6 PUFAs to n-3 PUFAs in pancreatic tissues would provide protection. Experiments were designed using n-3 fatty acid desaturase (Fat-1) transgenic mice, which can convert n-6 PUFA to n-3 FAs endogenously, to determine the impact of n-3 PUFAs on pancreatic intraepithelial neoplasms (PanINs) and their progression to pancreatic ductal adenocarcinoma (PDAC). Six-weekold female p48Cre/+-LSL-KrasG12D/+ andcompoundFat-1-p48Cre/+-LSL-KrasG12D/+ mice were fed (AIN-76A) diets containing 10% safflower oil for 35 weeks. Pancreata were evaluated histopathologically for PanINs and PDAC. Results showed a dramatic reduction in incidence of PDAC (84%; P < .02) in Fat-1-p48Cre/+-LSL-KrasG12D/+ mice compared to p48Cre/+-LSL-KrasG12D/+ mice. Importantly, significant reductions of pancreatic ducts with carcinoma (90%; P < .0001) and PanIN 3 (∼50%; P < .001) lesions were observed in the compound transgenic mice. The levels of n-3 PUFA were much higher (>85%; P < .05–0.01) in pancreas of compound transgenic mice than in those of p48Cre/+-LSL-KrasG12D/+ mice. Molecular analysis of the pancreas showed a significant down-regulation of proliferating cell nuclear antigen, cyclooxygenase-2, 5-lipoxygenase (5-LOX), 5-LOX-activating protein, Bcl-2, and cyclin D1 expression levels in Fat-1-p48Cre/+-LSL-KrasG12D/+ mice compared to p48Cre/+-LSL-KrasG12D/+ mice. These data highlight the promise of dietary n-3 FAs for chemoprevention of pancreatic cancer in high-risk individuals. PMID:23308056

  16. Stress-Induced Anxiety- and Depressive-Like Phenotype Associated with Transient Reduction in Neurogenesis in Adult Nestin-CreERT2/Diphtheria Toxin Fragment A Transgenic Mice.

    PubMed

    Yun, Sanghee; Donovan, Michael H; Ross, Michele N; Richardson, Devon R; Reister, Robin; Farnbauch, Laure A; Fischer, Stephanie J; Riethmacher, Dieter; Gershenfeld, Howard K; Lagace, Diane C; Eisch, Amelia J

    2016-01-01

    Depression and anxiety involve hippocampal dysfunction, but the specific relationship between these mood disorders and adult hippocampal dentate gyrus neurogenesis remains unclear. In both humans with MDD and rodent models of depression, administration of antidepressants increases DG progenitor and granule cell number, yet rodents with induced ablation of DG neurogenesis typically do not demonstrate depressive- or anxiety-like behaviors. The conflicting data may be explained by the varied duration and degree to which adult neurogenesis is reduced in different rodent neurogenesis ablation models. In order to test this hypothesis we examined how a transient-rather than permanent-inducible reduction in neurogenesis would alter depressive- and anxiety-like behaviors. Transgenic Nestin-CreERT2/floxed diphtheria toxin fragment A (DTA) mice (Cre+DTA+) and littermates (Cre+DTA-; control) were given tamoxifen (TAM) to induce recombination and decrease nestin-expressing stem cells and their progeny. The decreased neurogenesis was transient: 12 days post-TAM Cre+DTA+ mice had fewer DG proliferating Ki67+ cells and fewer DCX+ neuroblasts/immature neurons relative to control, but 30 days post-TAM Cre+DTA+ mice had the same DCX+ cell number as control. This ability of DG neurogenesis to recover after partial ablation also correlated with changes in behavior. Relative to control, Cre+DTA+ mice tested between 12-30 days post-TAM displayed indices of a stress-induced anxiety phenotype-longer latency to consume highly palatable food in the unfamiliar cage in the novelty-induced hypophagia test, and a depression phenotype-longer time of immobility in the tail suspension test, but Cre+DTA+ mice tested after 30 days post-TAM did not. These findings suggest a functional association between adult neurogenesis and stress induced anxiety- and depressive-like behaviors, where induced reduction in DCX+ cells at the time of behavioral testing is coupled with stress-induced anxiety and a

  17. Endogenous n-3 polyunsaturated fatty acids delay progression of pancreatic ductal adenocarcinoma in Fat-1-p48(Cre/+)-LSL-Kras(G12D/+) mice.

    PubMed

    Mohammed, Altaf; Janakiram, Naveena B; Brewer, Misty; Duff, Ashley; Lightfoot, Stan; Brush, Richard S; Anderson, Robert E; Rao, Chinthalapally V

    2012-12-01

    Preclinical studies suggest that diets rich in omega-3 polyunsaturated fatty acids (n-3 PUFAs) may be beneficial for prevention of pancreatic cancer. Nutritional intervention studies are often complex, and there is no clear evidence, without potential confounding factors, on whether conversion of n-6 PUFAs to n-3 PUFAs in pancreatic tissues would provide protection. Experiments were designed using n-3 fatty acid desaturase (Fat-1) transgenic mice, which can convert n-6 PUFA to n-3 FAs endogenously, to determine the impact of n-3 PUFAs on pancreatic intraepithelial neoplasms (PanINs) and their progression to pancreatic ductal adenocarcinoma (PDAC). Six-week-old female p48(Cre/+)-LSL-Kras(G12D/+) and compound Fat-1-p48(Cre/+)-LSL-Kras(G12D/+) mice were fed (AIN-76A) diets containing 10% safflower oil for 35 weeks. Pancreata were evaluated histopathologically for PanINs and PDAC. Results showed a dramatic reduction in incidence of PDAC (84%; P < .02) in Fat-1-p48(Cre/+)-LSL-Kras(G12D/+) mice compared to p48(Cre/+)-LSL-Kras(G12D/+) mice. Importantly, significant reductions of pancreatic ducts with carcinoma (90%; P < .0001) and PanIN 3 (~50%; P < .001) lesions were observed in the compound transgenic mice. The levels of n-3 PUFA were much higher (>85%; P < .05-0.01) in pancreas of compound transgenic mice than in those of p48(Cre/+)-LSL-Kras(G12D/+) mice. Molecular analysis of the pancreas showed a significant down-regulation of proliferating cell nuclear antigen, cyclooxygenase-2, 5-lipoxygenase (5-LOX), 5-LOX-activating protein, Bcl-2, and cyclin D1 expression levels in Fat-1-p48(Cre/+)-LSL-Kras(G12D/+) mice compared to p48(Cre/+)-LSL-Kras(G12D/+) mice. These data highlight the promise of dietary n-3 FAs for chemoprevention of pancreatic cancer in high-risk individuals. PMID:23308056

  18. A Novel Mgp-Cre Knock-In Mouse Reveals an Anticalcification/Antistiffness Candidate Gene in the Trabecular Meshwork and Peripapillary Scleral Region

    PubMed Central

    Borrás, Teresa; Smith, Matthew H.; Buie, LaKisha K.

    2015-01-01

    Purpose. Soft tissue calcification is a pathological condition. Matrix Gla (MGP) is a potent mineralization inhibitor secreted by cartilage chondrocytes and arteries' vascular smooth muscle cells. Mgp knock-out mice die at 6 weeks due to massive arterial calcification. Arterial calcification results in arterial stiffness and higher systolic blood pressure. Intriguingly, MGP was highly abundant in trabecular meshwork (TM). Because tissue stiffness is relevant to glaucoma, we investigated which additional eye tissues use Mgp's function using knock-in mice. Methods. An Mgp–Cre-recombinase coding sequence (Cre) knock-in mouse, containing Mgp DNA plus an internal ribosomal entry site (IRES)-Cre-cassette was generated by homologous recombination. Founders were crossed with Cre-mediated reporter mouse R26R-lacZ. Their offspring expresses lacZ where Mgp is transcribed. Eyes from MgpCre/+;R26RlacZ/+ (Mgp-lacZ knock-in) and controls, 1 to 8 months were assayed for β-gal enzyme histochemistry. Results. As expected, Mgp-lacZ knock-in's TM was intensely blue. In addition, this mouse revealed high specific expression in the sclera, particularly in the peripapillary scleral region (ppSC). Ciliary muscle and sclera above the TM were also positive. Scleral staining was located immediately underneath the choroid (chondrocyte layer), began midsclera and was remarkably high in the ppSC. Cornea, iris, lens, ciliary body, and retina were negative. All mice exhibited similar staining patterns. All controls were negative. Conclusions. Matrix Gla's restricted expression to glaucoma-associated tissues from anterior and posterior segments suggests its involvement in the development of the disease. Matrix Gla's anticalcification/antistiffness properties in the vascular tissue, together with its high TM and ppCS expression, place this gene as a strong candidate for TM's softness and sclera's stiffness regulation in glaucoma. PMID:25711639

  19. Defining the genome-wide role of CRE1 during carbon catabolite repression in Trichoderma reesei using RNA-Seq analysis.

    PubMed

    Antoniêto, Amanda Cristina Campos; dos Santos Castro, Lílian; Silva-Rocha, Rafael; Persinoti, Gabriela Felix; Silva, Roberto Nascimento

    2014-12-01

    The ascomycete Trichoderma reesei is one of the most well-studied cellulolytic fungi and is widely used by the biotechnology industry in the production of second generation bioethanol. The carbon catabolite repression (CCR) mechanism adopted by T. reesei is mediated by the transcription factor CRE1. CCR represses genes related to cellulase production when a carbon source is readily available in the medium. Using RNA sequencing, we investigated CCR during the synthesis of cellulases, comparing the T. reesei Δcre1 mutant strain with its parental strain, QM9414. Of 9129 genes in the T. reesei genome, 268 genes were upregulated and 85 were downregulated in the presence of cellulose (Avicel). In addition, 251 genes were upregulated and 230 were downregulated in the presence of a high concentration of glucose. Genes encoding cellulolytic enzymes and transcription factors and genes related to the transport of nutrients and oxidative metabolism were also targets of CCR, mediated by CRE1 in a carbon source-dependent manner. Our results also suggested that CRE1 regulates the expression of genes related to the use of copper and iron as final electron acceptors or as cofactors of enzymes that participate in biomass degradation. As a result, the final effect of CRE1-mediated transcriptional regulation is to modulate the access of cellulolytic enzymes to cellulose polymers or blocks the entry of cellulase inducers into the cell, depending on the glucose content in the medium. These results will contribute to a better understanding of the mechanism of carbon catabolite repression in T. reesei, thereby enhancing its application in several biotechnology fields. PMID:25459535

  20. Self-excision of the antibiotic resistance gene nptII using a heat inducible Cre-loxP system from transgenic potato.

    PubMed

    Cuellar, Wilmer; Gaudin, Amélie; Solórzano, Dennis; Casas, Armando; Nopo, Luis; Chudalayandi, Prakash; Medrano, Giuliana; Kreuze, Jan; Ghislain, Marc

    2006-09-01

    Resistance to antibiotics mediated by selectable marker genes remains a powerful selection tool for transgenic event production. However, regulatory agencies and consumer concerns favor these to be eliminated from food crops. Several excision systems exist but none have been optimized or shown to be functional for clonally propagated crops. The excision of the nptII gene conferring resistance to kanamycin has been achieved here using a gene construct based on a heat-inducible cre gene producing a recombinase that eliminates cre and nptII genes flanked by two loxP sites. First-generation regenerants with the Cre-loxP system were obtained by selection on kanamycin media. Following a heat treatment, second generation regenerants were screened for excision by PCR using nptII, cre, and T-DNA borders primers. Excision efficiency appeared to be at 4.7% depending on the heat treatment. The footprint of the excision was shown by sequencing between T-DNA borders to correspond to a perfect recombination event. Selectable marker-free sprouts were also obtained from tubers of transgenic events when submitted to similar heat treatment at 4% frequency. Spontaneous excision was not observed out of 196 regenerants from untreated transgenic explants. Biosafety concerns are minimized because the expression of cre gene driven by the hsp70 promoter of Drosophila melanogaster was remarkably low even under heat activation and no functional loxP site were found in published Solanum sequence database. A new plant transformation vector pCIP54/55 was developed including a multiple cloning site and the self-excision system which should be a useful tool not only for marker genes in potato but for any gene or sequence removal in any plant. PMID:16912912

  1. A CRE/AP-1-Like Motif Is Essential for Induced Syncytin-2 Expression and Fusion in Human Trophoblast-Like Model

    PubMed Central

    Vargas, Amandine; Rassart, Éric; Barbeau, Benoit

    2015-01-01

    Syncytin-2 is encoded by the envelope gene of Endogenous Retrovirus-FRD (ERVFRD-1) and plays a critical role in fusion of placental trophoblasts leading to the formation of the multinucleated syncytiotrophoblast. Its expression is consequently regulated in a strict manner. In the present study, we have identified a forskolin-responsive region located between positions -300 to -150 in the Syncytin-2 promoter region. This 150 bp region in the context of a minimal promoter mediated an 80-fold induction of promoter activity following forskolin stimulation. EMSA analyses with competition experiments with nuclear extracts from forskolin-stimulated BeWo cells demonstrated that the -211 to -177 region specifically bound two forskolin-induced complexes, one of them containing a CRE/AP-1-like motif. Site-directed mutagenesis of the CRE/AP-1 binding site in the context of the Syncytin-2 promoter or a heterologous promoter showed that this motif was mostly essential for forskolin-induced promoter activity. Transfection experiments with dominant negative mutants and constitutively activated CREB expression vectors in addition to Chromatin Immunoprecipitation suggested that a CREB family member, CREB2 was binding and acting through the CRE/AP-1 motif. We further demonstrated the binding of JunD to this same motif. Similar to forskolin and soluble cAMP, CREB2 and JunD overexpression induced Syncytin-2 promoter activity in a CRE/AP-1-dependent manner and Syncytin-2 expression. In addition, BeWo cell fusion was induced by both CREB2 and JunD overexpression, while being repressed following silencing of either gene. These results thereby demonstrate that induced expression of Syncytin-2 is highly dependent on the interaction of bZIP-containing transcription factors to a CRE/AP-1 motif and that this element is important for the regulation of Syncytin-2 expression, which results in the formation of the peripheral syncytiotrophoblast layer. PMID:25781974

  2. A Cre-lox approach for transient transgene expression in neural precursor cells and long-term tracking of their progeny in vitro and in vivo

    PubMed Central

    Geoffroy, Cédric G; Raineteau, Olivier

    2007-01-01

    Background Neural precursor cells (NPCs) can be isolated from various regions of the postnatal central nervous system (CNS). Manipulation of gene expression in these cells offers a promising strategy to manipulate their fate both in vitro and in vivo. In this study, we developed a technique that allows the transient manipulation of single/multiple gene expression in NPCs in vitro, and the long-term tracking of their progeny both in vitro and in vivo. Results In order to combine the advantages of transient transfection with the long-term tracking of the transfected cells progeny, we developed a new approach based on the cre-lox technology. We first established a fast and reliable protocol to isolate and culture NPCs as monolayer, from the spinal cord of neonatal transgenic Rosa26-YFP cre-reporter mice. These cells could be reliably transfected with single/multiple plasmids by nucleofection. Nucleofection with mono- or bicistronic plasmids containing the Cre recombinase gene resulted in efficient recombination and the long-term expression of the YFP-reporter gene. The transient cre-expression was non-toxic for the transfected cells and did not alter their intrinsic properties. Finally, we demonstrated that cre-transfected cells could be transplanted into the adult brain, where they maintained YFP expression permitting long-term tracking of their migration and differentiation. Conclusion This approach allows single/multiple genes to be manipulated in NPCs, while at the same time allowing long-term tracking of the transfected cells progeny to be analyzed both in vitro and in vivo. PMID:17504531

  3. Up, Up & Away. Strategic Campaigns.

    ERIC Educational Resources Information Center

    O'Shea, Catherine L.

    1999-01-01

    Uses a ballooning analogy to offer nine suggestions for universities conducting major fund-raising campaigns: study the map and choose a direction; test the prevailing winds; choose and train your crew; gear up for the journey; stay on course; make every bit count; change course as needed; and capitalize on your successful landing. (DB)

  4. African American Single Mothers Raising Sons: Implications for Family Therapy

    ERIC Educational Resources Information Center

    Gantt, Ann L.; Greif, Geoffrey L.

    2009-01-01

    Being raised by a single mother is one factor that has been suggested as contributing to the plight of African American males. Yet few studies have focused specifically on African American single mothers' experiences with raising sons. This qualitative study explored the following questions: (1) What are the experiences of African American single…

  5. An Introduction to Fund Raising: The Newcomers' Guide to Development.

    ERIC Educational Resources Information Center

    Faust, Paula J., Ed.

    A comprehensive introduction to all phases of college fund-raising is presented in 14 chapters. Attention is directed to the philosophy behind development, development's role in institutional advancement, and the personal qualities that lead to success in fund-raising. Basic principles also are covered for the following: annual giving, the capital…

  6. Consciousness Raising and Christian Worship as Small Group Communication.

    ERIC Educational Resources Information Center

    Burns, Gary

    Consciousness raising movements and charismatic Christian worship display an extraordinary degree of rhetorical similarity. This four-part paper outlines the likenesses of the two groups, indicates where they differ, and focuses on the social and political dimensions of consciousness raising. The first section lists the following similarities…

  7. Grandparents Raising Their Grandchildren: A Comparative Study of Depression

    ERIC Educational Resources Information Center

    Strutton, Joan N.

    2010-01-01

    There has been a steady increase in the United States in recent decades of grandparents raising their grandchildren. The aim of this study was to determine if depression levels of grandparents raising their grandchildren and depression levels of traditional grandparents differ. Additionally, the extent of the relationship to depression scores by…

  8. Phonological Consciousness Raising Tasks for the ESP Classroom.

    ERIC Educational Resources Information Center

    Sterlacci, Peter

    A discussion of pronunciation instruction in English for business communication focuses on raising learners' awareness of how English is spoken by various language groups around the world. It is argued that phonological consciousness-raising is an effective approach for limiting breakdowns in communication. The approach assumes that developing…

  9. 47 CFR 25.282 - Orbit raising maneuvers.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 47 Telecommunication 2 2010-10-01 2010-10-01 false Orbit raising maneuvers. 25.282 Section 25.282... Technical Operations § 25.282 Orbit raising maneuvers. A space station authorized to operate in the geostationary satellite orbit under this part is also authorized to transmit in connection with...

  10. 47 CFR 25.282 - Orbit raising maneuvers.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 47 Telecommunication 2 2012-10-01 2012-10-01 false Orbit raising maneuvers. 25.282 Section 25.282... Technical Operations § 25.282 Orbit raising maneuvers. A space station authorized to operate in the geostationary satellite orbit under this part is also authorized to transmit in connection with...

  11. 47 CFR 25.282 - Orbit raising maneuvers.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 47 Telecommunication 2 2013-10-01 2013-10-01 false Orbit raising maneuvers. 25.282 Section 25.282... Technical Operations § 25.282 Orbit raising maneuvers. A space station authorized to operate in the geostationary satellite orbit under this part is also authorized to transmit in connection with...

  12. 47 CFR 25.282 - Orbit raising maneuvers.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 47 Telecommunication 2 2011-10-01 2011-10-01 false Orbit raising maneuvers. 25.282 Section 25.282... Technical Operations § 25.282 Orbit raising maneuvers. A space station authorized to operate in the geostationary satellite orbit under this part is also authorized to transmit in connection with...

  13. 47 CFR 25.282 - Orbit raising maneuvers.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 47 Telecommunication 2 2014-10-01 2014-10-01 false Orbit raising maneuvers. 25.282 Section 25.282... Technical Operations § 25.282 Orbit raising maneuvers. A space station authorized to operate in the geostationary satellite orbit under this part is also authorized to transmit in connection with...

  14. 33 CFR 401.39-1 - Raising fenders.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Raising fenders. 401.39-1 Section 401.39-1 Navigation and Navigable Waters SAINT LAWRENCE SEAWAY DEVELOPMENT CORPORATION, DEPARTMENT OF TRANSPORTATION SEAWAY REGULATIONS AND RULES Regulations Seaway Navigation § 401.39-1 Raising fenders....

  15. HUNTER 10, TRUFLO GONDOLA WEIGHTS AND JACKET. PIVOTING ARMS RAISE ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    HUNTER 10, TRUFLO GONDOLA WEIGHTS AND JACKET. PIVOTING ARMS RAISE THE JACKET AND WEIGHTS. A PNEUMATIC SHOVEL PUSHES THE MORE RECENTLY POURED MOLD INTO THE COOLED MOLD, FORCING THE LATTER ONTO THE VIBRATING SHAKEOUT CONVEYOR AND LEAVING A SPACE UNDER THE RAISED JACKET AND WEIGHTS FOR A FRESHLY MADE UNPOURED MOLD. - Southern Ductile Casting Company, Casting, 2217 Carolina Avenue, Bessemer, Jefferson County, AL

  16. Fund Raising in California School Districts. A Discussion Paper.

    ERIC Educational Resources Information Center

    Allen, Thomas W.; Hughes, K. Scott

    The results of a 1982 survey of California school districts concerning their efforts at private fundraising are presented here. It was found that 123 (or 61 percent) of responding districts either have programs or are planning programs. Although six districts raised more than $100,000 in 1981-82, the majority raised less that $20,000. Of the 66…

  17. Constructing an Engineering Model for Raising an Egyptian Obelisk

    ERIC Educational Resources Information Center

    Beck, Charles R.

    2009-01-01

    One of the greatest mysteries of ancient times is how the Egyptians managed to raise huge obelisks using very simple technology. This remarkable task has puzzled engineers for thousand of years. After failing to raise an obelisk with simple machines, such as levers and pulleys, a team of modern engineers solved the mystery using a sandpit and the…

  18. Strategic planning decision making using fuzzy SWOT-TOPSIS with reliability factor

    NASA Astrophysics Data System (ADS)

    Mohamad, Daud; Afandi, Nur Syamimi; Kamis, Nor Hanimah

    2015-10-01

    Strategic planning is a process of decision making and action for long-term activities in an organization. The Strengths, Weaknesses, Opportunities and Threats (SWOT) analysis has been commonly used to help organizations in strategizing their future direction by analyzing internal and external environment. However, SWOT analysis has some limitations as it is unable to prioritize appropriately the multiple alternative strategic decisions. Some efforts have been made to solve this problem by incorporating Multi Criteria Decision Making (MCDM) methods. Nevertheless, another important aspect has raised concerns on obtaining the decision that is the reliability of the information. Decision makers evaluate differently depending on their level of confidence or sureness in the evaluation. This study proposes a decision making procedure for strategic planning using SWOT-TOPSIS method by incorporating the reliability factor of the evaluation based on Z-number. An example using a local authority in the east coast of Malaysia is illustrated to determine the strategic options ranking and to prioritize factors in each SWOT category.

  19. NERSC Strategic Implementation Plan 2002-2006

    SciTech Connect

    Kramer, William; Bethel, Wes; Craw, James; Draney, Brent; Fortney, William; Gorda, Brend; Harris, William; Meyer, Nancy; Ng, Esmond; Verdier, Francesca; Walter, Howard; Welcome, Tammy

    2002-09-01

    This strategic proposal presents NERSC's vision for its activities and new directions over the next five years. NERSC's continuing commitment to providing high-end systems and comprehensive scientific support for its users will be enhanced, and these activities will be augmented by two new strategic thrusts: support for Scientific Challenge Teams and deployment of a Unified Science Environment. The proposal is in two volumes, the Strategic Plan and the Implementation Plan.

  20. Reflections on strategic nurse leadership.

    PubMed

    White, Jean

    2012-10-01

    This paper sets out some personal reflections by the Chief Nursing Officer for Wales on the challenges facing nurses and midwives as they undertake strategic leadership roles in NHS organisations. The paper looks at the national approach taken in Wales where behavioural competencies for executive nurse directors have been implemented. It considers the implications of the breadth of responsibilities executive nurse directors have and the importance of developing and supporting middle grade nurse managers and clinical directors. It concludes by looking at who is responsible for care within an organisation. PMID:23050616

  1. Strategic workshops on cancer nanotechnology.

    PubMed

    Nagahara, Larry A; Lee, Jerry S H; Molnar, Linda K; Panaro, Nicholas J; Farrell, Dorothy; Ptak, Krzysztof; Alper, Joseph; Grodzinski, Piotr

    2010-06-01

    Nanotechnology offers the potential for new approaches to detecting, treating, and preventing cancer. To determine the current status of the cancer nanotechnology field and the optimal path forward, the National Cancer Institute's Alliance for Nanotechnology in Cancer held three strategic workshops, covering the areas of in vitro diagnostics and prevention, therapy and post-treatment, and in vivo diagnosis and imaging. At each of these meetings, a wide range of experts from academia, industry, the nonprofit sector, and the U.S. government discussed opportunities in the field of cancer nanotechnology and barriers to its implementation. PMID:20460532

  2. Strategic defense initiative: Folly or future

    SciTech Connect

    Haley, P.E.; Merritt, J.

    1986-01-01

    This collection of analyses is a guide through the maze of claims and criticisms about ''Star Wars,'' the controversial effort of the Reagan administration to reorient United States nuclear strategy to strategic defense. The text starts with an introduction by the editors followed by individual chapters outlining the strategic defense initiative as originally conceived and subsequently modified by the Reagan administration; the arguments for and against the plan's strategic and technical feasibility; and assessments of the harmful and constructive effects of strategic defense on U.S.-Soviet and U.S.-allied relations.

  3. Excellence in Educational Fund Raising at America's Community Colleges and a Key Resources Guide for Educational Fund Raising.

    ERIC Educational Resources Information Center

    Ryan, G. Jeremiah

    Designed to encourage and inform community college efforts to secure private financial support, this literature review and resource guide examine the current status of fund raising at community colleges and list pertinent information sources. After introductory comments advocate increased community college involvement in fund raising, the paper…

  4. 77 FR 54615 - Strategic Management Program; Fiscal Year 2013-2016 Strategic Plan

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-05

    ... From the Federal Register Online via the Government Publishing Office NATIONAL TRANSPORTATION SAFETY BOARD Strategic Management Program; Fiscal Year 2013-2016 Strategic Plan AGENCY: National....ntsb.gov/doclib/agency_reports/Strategic-Plan_2013-2016.pdf . DATES: Parties should submit comments...

  5. Enhancement by lithium of cAMP-induced CRE/CREB-directed gene transcription conferred by TORC on the CREB basic leucine zipper domain

    PubMed Central

    Böer, Ulrike; Eglins, Julia; Krause, Doris; Schnell, Susanne; Schöfl, Christof; Knepel, Willhart

    2007-01-01

    The molecular mechanism of the action of lithium salts in the treatment of bipolar disorder is not well understood. As their therapeutic action requires chronic treatment, adaptive neuronal processes are suggested to be involved. The molecular basis of this are changes in gene expression regulated by transcription factors such as CREB (cAMP-response-element-binding protein). CREB contains a transactivation domain, in which Ser119 is phosphorylated upon activation, and a bZip (basic leucine zipper domain). The bZip is involved in CREB dimerization and DNA-binding, but also contributes to CREB transactivation by recruiting the coactivator TORC (transducer of regulated CREB). In the present study, the effect of lithium on CRE (cAMP response element)/CREB-directed gene transcription was investigated. Electrically excitable cells were transfected with CRE/CREB-driven luciferase reporter genes. LiCl (6 mM or higher) induced an up to 4.7-fold increase in 8-bromo-cAMP-stimulated CRE/CREB-directed transcription. This increase was not due to enhanced Ser119 phosphorylation or DNA-binding of CREB. Also, the known targets inositol monophosphatase and GSK3β (glycogen-synthase-kinase 3β) were not involved as specific GSK3β inhibitors and inositol replenishment did not mimic and abolish respectively the effect of lithium. However, lithium no longer enhanced CREB activity when the CREB-bZip was deleted or the TORC-binding site inside the CREB-bZip was specifically mutated (CREB-R300A). Otherwise, TORC overexpression conferred lithium responsiveness on CREB-bZip or the CRE-containing truncated rat somatostatin promoter. This indicates that lithium enhances cAMP-induced CRE/CREB-directed transcription, conferred by TORC on the CREB-bZip. We thus support the hypothesis that lithium salts modulate CRE/CREB-dependent gene transcription and suggest the CREB coactivator TORC as a new molecular target of lithium. PMID:17696880

  6. v-src induction of the TIS10/PGS2 prostaglandin synthase gene is mediated by an ATF/CRE transcription response element.

    PubMed Central

    Xie, W; Fletcher, B S; Andersen, R D; Herschman, H R

    1994-01-01

    We recently reported the cloning of a mitogen-inducible prostaglandin synthase gene, TIS10/PGS2. In addition to growth factors and tumor promoters, the v-src oncogene induces TIS10/PGS2 expression in 3T3 cells. Deletion analysis, using luciferase reporters, identifies a region between -80 and -40 nucleotides 5' of the TIS10/PGS2 transcription start site that mediates pp60v-src induction in 3T3 cells. This region contains the sequence CGTCACGTG, which includes overlapping ATF/CRE (CGTCA) and E-box (CACGTG) sequences. Gel shift-oligonucleotide competition experiments with nuclear extracts from cells stably transfected with a temperature-sensitive v-src gene demonstrate that the CGTCACGTG sequence can bind proteins at both the ATF/CRE and E-box sequences. Dominant-negative CREB and Myc proteins that bind DNA, but do not transactivate, block v-src induction of a luciferase reporter driven by the first 80 nucleotides of the TIS10/PGS2 promoter. Mutational analysis distinguishes which TIS10/PGS2 cis-acting element mediates pp60v-src induction. E-box mutation has no effect on the fold induction in response to pp60v-src. In contrast, ATF/CRE mutation attenuates the pp60v-src response. Antibody supershift and methylation interference experiments demonstrate that CREB and at least one other ATF transcription factor in these extracts bind to the TIS10/PGS2 ATF/CRE element. Expression of a dominant-negative ras gene also blocks TIS10/PGS2 induction by v-src. Our data suggest that Ras mediates pp60v-src activation of an ATF transcription factor, leading to induced TIS10/PGS2 expression via the ATF/CRE element of the TIS10/PGS2 promoter. This is the first description of v-src activation of gene expression via an ATF/CRE element. Images PMID:7935375

  7. Strategic Petroleum Reserve annual report for calendar year 1998

    SciTech Connect

    1998-12-31

    The Strategic Petroleum Reserve was established in 1975 as an emergency response to the 1973 Arab oil embargo. It is authorized by the Energy Policy and Conservation Act (EPCA), and by the comprehensive energy plans of all Administrations since 1975, in recognition of the long-term dependence of the US on imported crude oil and petroleum products. Section 165 of EPCA requires the Secretary of Energy to submit an Annual Report to the President and the Congress. On May 13, 1998, the Department published a Statement of Administration Policy which reaffirmed its commitment to maintain a Government-owned and controlled, centrally located Strategic Petroleum Reserve of crude oil. The Reserve is to be used solely for responding to the types of severe oil supply interruptions presently contemplated in EPCA. Over the past twenty years, the Reserve has grown as large as 592 million barrels--a peak reached in 1994. From 1994 to 1996, nearly 28 million barrels were sold to raise revenues for the U S Treasury. As of December 31, 1998, the crude oil inventory was 561,108,127 barrels which equated to 60 days of net oil imports during 1998. The US now relies on a combination of both the Reserve and private stocks to meet its oil storage obligations to the International Energy Agency.

  8. Textpresso Site-Specific Recombinases: a text-mining server for the recombinase literature including Cre mice and conditional alleles

    PubMed Central

    Urbanski, William M.; Condie, Brian G.

    2016-01-01

    Textpresso Site Specific Recombinases (http://ssrc.genetics.uga.edu/) is a text-mining web server for searching a database of over 9000 full-text publications. The papers and abstracts in this database represent a wide range of topics related to site-specific recombinase (SSR) research tools. Included in the database are most of the papers that report the characterization or use of mouse strains that express Cre recombinase as well as papers that describe or analyze mouse lines that carry conditional (floxed) alleles or SSR-activated transgenes/knockins. The database also includes reports describing SSR-based cloning methods such as the Gateway or the Creator systems, papers reporting the development or use of SSR-based tools in systems such as Drosophila, bacteria, parasites, stem cells, yeast, plants, zebrafish and Xenopus as well as publications that describe the biochemistry, genetics or molecular structure of the SSRs themselves. Textpresso Site Specific Recombinases is the only comprehensive text-mining resource available for the literature describing the biology and technical applications of site-specific recombinases. PMID:19882667

  9. Ar-Ar Ages of Nakhlites Y000593, NWA998, and Nakhla and CRE Ages of NWA998

    NASA Technical Reports Server (NTRS)

    Garrison, D. H.; Bogard, D. D.

    2005-01-01

    The seven known Martian nakhlites are Nakhla, Lafayette, Governador Valadares, and four recent finds from hot and cold deserts: MIL03346 from the Transantarctic Mountains, a paired group from the Yamato Mountains (Y000593, Y000749, and Y000802, and two from Morocco (NWA998 and NWA817. Radiometric ages (Sm-Nd, Rb-Sr, U-Pb, and Ar-Ar) for the first three nakhlites, along with Chassigny, fall in the range of 1.19-1.37 Gyr and may suggest a common formation age. These meteorites also show very similar cosmic-ray (space) exposure ages, suggesting a single ejection event from Mars. The ages for nakhlites are different from those of Martian shergottites, whose radiometric ages vary by nearly a factor of three (approx. 165-475 Myr) and whose space exposure ages vary over a factor of approx. 20. Shergottite ages suggest that multiple locations on the Martian surface have been sampled, whereas nakhlite data imply sampling of only one Mars surface location. Because older Martian surfaces are expected to be more abundant, it seems surprising that all nakhlites would represent only one Martian impact event. To address this issue, we are measuring the (39)Ar-(40)Ar ages of Y-000593, NWA-998, Nakhla, and MIL-03346, and the space (CRE) exposure age of NWA998. Additional information is included in the original extended abstract.

  10. A novel Cre recombinase reporter mouse strain facilitates selective and efficient infection of primary immune cells with adenoviral vectors.

    PubMed

    Heger, Klaus; Kober, Maike; Rieß, David; Drees, Christoph; de Vries, Ingrid; Bertossi, Arianna; Roers, Axel; Sixt, Michael; Schmidt-Supprian, Marc

    2015-06-01

    Replication-deficient recombinant adenoviruses are potent vectors for the efficient transient expression of exogenous genes in resting immune cells. However, most leukocytes are refractory to efficient adenoviral transduction as they lack expression of the coxsackie/adenovirus receptor (CAR). To circumvent this obstacle, we generated the R26/CAG-CARΔ1(StopF) (where R26 is ROSA26 and CAG is CMV early enhancer/chicken β actin promoter) knock-in mouse line. This strain allows monitoring of in situ Cre recombinase activity through expression of CARΔ1. Simultaneously, CARΔ1 expression permits selective and highly efficient adenoviral transduction of immune cell populations, such as mast cells or T cells, directly ex vivo in bulk cultures without prior cell purification or activation. Furthermore, we show that CARΔ1 expression dramatically improves adenoviral infection of in vitro differentiated conventional and plasmacytoid dendritic cells (DCs), basophils, mast cells, as well as Hoxb8-immortalized hematopoietic progenitor cells. This novel dual function mouse strain will hence be a valuable tool to rapidly dissect the function of specific genes in leukocyte physiology. PMID:25787118

  11. Protocols for Analyzing the Role of Paneth Cells in Regenerating the Murine Intestine using Conditional Cre-lox Mouse Models

    PubMed Central

    Parry, Lee; Young, Madeleine; El Marjou, Fatima; Clarke, Alan Richard

    2015-01-01

    The epithelial surface of the mammalian intestine is a dynamic tissue that renews every 3 - 7 days. Understanding this renewal process identified a population of rapidly cycling intestinal stem cells (ISCs) characterized by their expression of the Lgr5 gene. These are supported by a quiescent stem cell population, marked by Bmi-1 expression, capable of replacing them in the event of injury. Investigating the interactions between these populations is crucial to understanding their roles in disease and cancer. The ISCs exist within crypts on the intestinal surface, these niches support the ISC in replenishing the epithelia. The interaction between active and quiescent ISCs likely involves other differentiated cells within the niche, as it has previously been demonstrated that the ‘‘stemness’’ of the Lgr5 ISC is closely tied to the presence of their neighboring Paneth cells. Using conditional cre-lox mouse models we tested the effect of deleting the majority of active ISCs in the presence or absence of the Paneth cells. Here we describe the techniques and analysis undertaken to characterize the intestine and demonstrate that the Paneth cells play a crucial role within the ISC niche in aiding recovery following substantial insult. PMID:26649885

  12. Building Bridges: Fund Raising for Deans, Faculty, and Development Officers.

    ERIC Educational Resources Information Center

    Murphy, Mary Kay, Ed.

    This publication offers 16 articles by higher education administrators, faculty and development officers on dean and faculty members roles in development work as well as the teamwork and synergy that develop in the best efforts. Following an introduction by George Keller, the articles are as follows: (1) "Institutional Strategic Planning: Setting…

  13. Collaborative Strategic Planning in Higher Education

    ERIC Educational Resources Information Center

    Sanaghan, Patrick

    2009-01-01

    This book outlines a simple, five-phase collaborative approach to strategic planning that has worked effectively on many campuses. Specifically, Collaborative Strategic Planning (CSP) refers to the disciplined and thoughtful process of meaningfully engaging relevant stakeholders in creating a shared future vision and goals for their institution.…

  14. Strategic Teaching: Some Concepts for Instructional Design.

    ERIC Educational Resources Information Center

    Jones, Beau Fly

    1988-01-01

    Discusses strategic teaching, a process for organizing and representing subject content, for helping students think about their own learning processes, for organizing skills instruction, and for managing classroom instruction. Provides an example of strategic instructional planning and a graphic that shows step-by-step procedures for the planning…

  15. Using Appropriate Tools Strategically for Instruction

    ERIC Educational Resources Information Center

    Sherman, Milan; Cayton, Charity

    2015-01-01

    Students' ability to use appropriate tools strategically is an important skill of mathematically proficient students (SMP 5, CCSSI 2010, p. 7). A parallel practice for teachers is using appropriate tools strategically for mathematics instruction. An important element of this practice is that the use of technology depends on the goals of…

  16. Using Strategic Planning To Improve Rural Schools.

    ERIC Educational Resources Information Center

    D'Amico, Joseph J.

    This paper describes the elements of strategic planning and applies them to educational reform and improvement. The paper also describes how a planning model, Strategic Planning for Educational Reform and Improvement (SPERI), was used by Rural Assistance Councils (RACs) in Pennsylvania and Delaware, with technical assistance from Research for…

  17. 24 CFR 91.415 - Strategic plan.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 24 Housing and Urban Development 1 2014-04-01 2014-04-01 false Strategic plan. 91.415 Section 91... Plan § 91.415 Strategic plan. Strategies and priority needs must be described in the consolidated plan... required to submit a nonhousing Community Development Plan; however, if the consortium includes...

  18. 24 CFR 91.415 - Strategic plan.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 24 Housing and Urban Development 1 2013-04-01 2013-04-01 false Strategic plan. 91.415 Section 91... Plan § 91.415 Strategic plan. Strategies and priority needs must be described in the consolidated plan... required to submit a nonhousing Community Development Plan; however, if the consortium includes...

  19. 24 CFR 91.415 - Strategic plan.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 24 Housing and Urban Development 1 2011-04-01 2011-04-01 false Strategic plan. 91.415 Section 91... Plan § 91.415 Strategic plan. Strategies and priority needs must be described in the consolidated plan... required to submit a nonhousing Community Development Plan; however, if the consortium includes...

  20. 24 CFR 91.215 - Strategic plan.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 24 Housing and Urban Development 1 2014-04-01 2014-04-01 false Strategic plan. 91.215 Section 91.215 Housing and Urban Development Office of the Secretary, Department of Housing and Urban Development CONSOLIDATED SUBMISSIONS FOR COMMUNITY PLANNING AND DEVELOPMENT PROGRAMS Local Governments; Contents of Consolidated Plan § 91.215 Strategic plan....