To clarify whether transduction efficiency and cell type specificity of self-complementary (sc) AAV5 vectors are similar to those of standard, single stranded AAV5 vectors in normal retina, one micro liter of scAAV5-smCBA-GFP vector ...

This study investigated whether repeated administration of recombinant adeno-associated virus type 5 (rAAV5) to the airways induces inflammatory processes in the lungs of BALB/c-mice, with mechanical and histologic changes. Saline was instilled intratracheally in the control group, and rAAV5-green fluorescence protein (GFP) (4x10(11)particles) in the virus ...

Lung infections with Pseudomonas aeruginosa and other pathogens in cystic fibrosis (CF) cause progressive airway obstruction and tissue damage, the predominant cause of morbidity and mortality in CF. We investigated whether a recombinant adeno-associated virus type 5 (AAV5) vector expressing murine interleukin (IL)-10 ...

Recombinant adeno-associated virus (AAV) vectors are promising tools for gene therapy. In spinal cord injury where extensive damage occurs, vectors with high diffusion and transduction abilities are required. We compared the diffusion capacity and transduction efficiency of AAV2 and AAV5 vectors using a mouse spinal cord injury model. ...

Abstract Intravascular delivery of adeno-associated virus (AAV) vector is commonly used for liver-directed gene therapy. In humans, the high prevalence of neutralizing antibodies to AAV-2 capsid and the wide cross-reactivity with other serotypes hamper vector transduction efficacy. Moreover, the safety of gene-based approaches depends on ...

Untargeted and uncontrolled gene delivery is a major cause of gene therapy failure. This study aimed to define efficient and safe tissue-selective targeted gene therapy approaches for delivering genes into keratocytes of the cornea in vivo using a normal or diseased rabbit model. New Zealand White rabbits, adeno-associated virus serotype 5 (AAV5), and a ...

Untargeted and uncontrolled gene delivery is a major cause of gene therapy failure. This study aimed to define efficient and safe tissue-selective targeted gene therapy approaches for delivering genes into keratocytes of the cornea in vivo using a normal or diseased rabbit model. New Zealand White rabbits, adeno-associated virus serotype 5 (AAV5), and a ...

Since transgenes were first cloned into recombinant adenoviruses almost 30 years ago, a variety of viral vectors have become important tools in genetic research. Viruses adeptly transport genetic material into eukaryotic cells, and replacing all or part of the viral genome with genes of interest or silencing sequences creates a method of gene expression modulation in which the ...

Purpose. We previously showed that subretinal delivery of AAV5 vectors containing murine GC1 cDNA driven by either photoreceptor-specific, (hGRK1) or ubiquitous (smCBA) promoters were capable of restoring cone-mediated function and visual behavior and preserving cone photoreceptors in the guanylatecylase 1 knockout (GC1KO) mouse for three months. Here, we compare therapy ...

Viral vector-mediated gene transfer has emerged as a powerful means to target transgene expression in the central nervous system. Here we characterized the efficacy of serotypes 1 and 5 recombinant adeno-associated virus (rAAV) vectors encoding green fluorescent protein (GFP) after stereotaxic delivery to the neonatal rat and minipig striatum. The ...

AbstractRecombinant adeno-associated viral vectors pseudotyped with serotype 5 and 8 capsids (AAV5 and AAV8) have been shown to be efficient gene transfer reagents for the liver. We have produced AAV5 and AAV8 vectors that express mouse short-chain acyl-CoA dehydrogenase ...

Abstract Recombinant adeno-associated virus 5 (rAAV5) represents a candidate vector with unique advantages for the treatment of hepatic disorders because of its narrow hepatic tropism. Noninvasive in vivo imaging of transgene expression provides an important tool with which to quantify the transduction efficiency, and duration and ...

BackgroundNeuronal transduction by adeno-associated viral (AAV) vectors has been demonstrated in cortex, brainstem, cerebellum, and sensory ganglia. Intrathecal delivery of AAV serotypes that transduce neurons in dorsal root ganglia (DRG) and spinal cord offers substantial opportunities to 1) further study mechanisms underlying chronic pain, and 2) develop novel gene-based ...

Adeno-associated virus (AAV) provides a promising platform for clinical treatment of neurological disorders owing to its established efficacy and lack of apparent pathogenicity. To use viral vectors in treating neurological disease, however, transduction must occur under neuropathological conditions. Previous studies in rodents have shown that AAV5 more efficiently transduces ...

SummaryTo test whether fast-acting, self complimentary(sc), AAV vector-mediated RPE65 expression prevents cone degeneration and/or restores cone function, two mouse lines were studied: the Rpe65- deficient rd12 mouse and the Rpe65- deficient, rhodpsin null (�i.e. cone function-only�) Rpe65^?/?::Rho^?/? mouse. ...

Selecting the most efficient recombinant adeno-associated virus (rAAV) serotype for airway gene therapy has been difficult due to cross-specific differences in tropism and immune response between humans and animal models. Chimpanzees�the closest surviving genetic relative of humans�provide a valuable opportunity to select the most effective serotypes for clinical trials in humans. However, ...

... Accession Number : ADA512050. Title : Emergence of a New Neotropical Malaria Vector Facilitated by Human Migration and Changes in Land Use. ...

To test whether fast-acting, self-complimentary (sc), adeno-associated virus-mediated RPE65 expression prevents cone degeneration and/or restores cone function, we studied two mouse lines: the Rpe65-deficient rd12 mouse and the Rpe65-deficient, rhodopsin null ('that is, cone function-only') Rpe65(-/-)::Rho(-/-) mouse. scAAV5 expressing RPE65 was injected ...

Differences between rodent and human airway cell biology have made it difficult to translate recombinant adeno-associated virus (rAAV)-mediated gene therapies to the lung for cystic fibrosis (CF). As new ferret and pig models for CF become available, knowledge about host cell/vector interactions in these species will become increasingly important for testing potential gene ...

Vectors derived from adeno-associated virus (AAV) are promising candidates for neural cell transduction in vivo because they are nonpathogenic and achieve long-term transduction in the central nervous system. AAV serotype 2 (AAV2) is the most widely used AAV vector in clinical trials based largely on its ability to transduce neural cells in the rodent and ...

The helper-dependent adeno-associated viruses (AAVs) have attracted great interest as vectors for gene therapy. Uptake and intracellular trafficking pathways of AAV are of importance, since they are often rate-limiting steps in infection. Here, we have investigated the entry of AAV type 5 (AAV5) in primary human embryo fibroblasts. At ...

BackgroundThe current method of delivering gene replacement to the posterior segment of the eye involves a three-port pars plana vitrectomy followed by injection of the agent through a 37-gauge cannula, which is potentially wrought with retinal complications. In this paper we investigate the safety and efficacy of delivering adeno-associated viral (AAV) vector to the ...

... 10. Linthicum KJ, 1988. A revision of the Argyritarsis Section of the subgenus Nyssorhynchus of Anopheles (Diptera: Culici- dae). ...

facilitates testing of the EMA. Tests such as frequency response, stability, and step response are discussed in more detail in the next section. ...

The language Vector C is a superset of the conventional (scalar) programming language C with extensions to facilitate vector processing. The design goals and a methodology for performing language extensions and the extended language features of Vector C are presented. The implementation of ...

PurposeWith the increasing number of retinal gene-based therapies and therapeutic constructs, in vitro bioassays characterizing vector transduction efficiency and quality are becoming increasingly important. Currently, in vitro assays quantifying vector transduction efficiency are performed predominantly for non-ocular tissues. A human retinal pigment ...

Globoid cell leukodystrophy (GLD) is a devastating lysosomal storage disease caused by deficiency of the enzyme galactocerebrosidase (GALC). Currently, there is no definite cure for GLD. Several attempts with CNS-directed gene therapy in twitcher mice (a murine model of GLD) demonstrated restricted expression of GALC activity in CNS and failure of therapeutic efficacy in cerebellum and spinal ...

Although recombinant adeno-associated virus (rAAV) has been widely used in lung gene therapy approaches, it remains unclear to what extent commonly used AAV serotypes transduce adult progenitors in the lung. In this study, we evaluated the life span and proliferative capacity of rAAV1-, 2-, and 5-transduced airway cells in mouse lung, using a LacZ-CRE reporter transgenic model and Cre-expressing ...

The development of Alzheimer's disease (AD) is closely connected with cholesterol metabolism. Cholesterol increases the production and deposition of amyloid-? (A?) peptides that result in the formation of amyloid plaques, a hallmark of the pathology. In the brain, cholesterol is synthesized in situ but cannot be degraded nor cross the blood�brain barrier. The major exportable form of brain ...

Rous sarcoma virus (RSV) can be used for the simple generation of high-titer replication-competent retroviral (RCR) vectors. Retroviruses undergo frequent genomic recombination, however, and vectors with reduced replication kinetics are rapidly overgrown by mutant forms. Vector design is hence critical to vector ...

In a region of northeastern Amazonia, we find a species previously of minor importance, Anopheles marajoara, to be the principal malaria vector. In a total of five collections during 1996-97 in three replicated sites near the city of Macapa, Amapa state, ...

... Wheatbelt may facilitate greater abundance of halotolerant mosquitoes, Aedes alboannulatus Macquart and Aedes camptorhynchus Thomson (a vector of Ross River virus [ ... ...

This chapter is devoted to a discussion of three topics: The discovery of the major etiologic agents of rickettsial disease, their vectors, and mechanisms of transmission; The major technological developments that have facilitated the study, control, and ...

... Issuances (current and cancelled), Joint Staff and ... Software Initiative (HPEC-SI) Program Facilitation of ... Vector Signal and Image Processing Library ...

... enclosing the plants in perforated plastic bags and fastening the bag around each pot using an elastic ... ...

A detector for the Superconducting Super Collider is outlined and its key parameters are fixed to facilitate the study of very heavy flavors and heavy vector bosons as well as the Pomeron. (LEW)

Research on the genetics of methane producing archaebacteria is directed at fundamental problems of mutant construction, gene transfer by transformation, and construction of vectors which facilitate gene transfer. From the thermophilic methanogen, Methano...

... culture by transfecting with human siat7e gene and ... Viruses as Bacterial Artificial Chromosomes to Facilitate ... simplex virus 2 bacterial artificial chromosome. ...

A novel algorithm facilitates the wind vector analysis of the HARLIE data. ... from exchange of axial angular momentum between the solid mantle and the core ...

Fifteen related ligation-independent cloning vectors were constructed for high-throughput cloning and purification of proteins. The vectors encode a TEV protease site for removal of tags that facilitate pro tein purification (his-tag) or improve solubility (MBP, GST). Specialized vectors allow coexpression and ...

SummaryFifteen related ligation-independent cloning vectors were constructed for high-throughput cloning and purification of proteins. The vectors encode a TEV protease site for removal of tags that facilitate protein purification (his-tag) or improve solubility (MBP, GST). Specialized vectors allow coexpression ...

Despite the extensive research efforts over the past 25 years that have focused on HIV, there is still no cure for AIDS. However, tremendous progress in the understanding of the structure and biology of the HIV virus led to the development of safe and potent HIV-based transgene delivery vectors. These genetic vehicles are referred to as lentiviral vectors. ...

The broad application of retroviral vectors for gene delivery is still hampered by the difficulty to reproducibly establish high vector producer cell lines generating sufficient amounts of highly concentrated virus vector preparations of high quality. To enhance the process for producing clinically relevant retroviral ...

The restricted host-cell range and low titer of retroviral vectors limit their use for stable gene transfer in eukaryotic cells. To overcome these limitations, we have produced murine leukemia virus-derived vectors in which the retroviral envelope glycoprotein has been completely replaced by the G glycoprotein of vesicular stomatitis virus. Such ...

The single-stranded genome of adeno-associated virus (AAV) vectors is one of the key factors leading to a slow-rising but long-term transgene expression kinetics. Previous molecular studies have established what is now considered as a textbook molecular model of AAV genomes with two copies of inverted-tandem repeats at either end. In this study, We profiled hundreds of ...

Recent developments in the use of viruses as vaccine vectors have been facilitated by a better understanding of viral biology. Advances occur as we gain greater insight into the interrelationship of viruses and the immune system. Viral-vector vaccines remain the best means to induce cellular immunity and are now showing promise for the ...

Despite their central role in disease transmission, relatively little is known of the molecular biology of arthropod vectors. Modern molecular approaches will undoubtedly provide considerable information about gene regulation and expression in vectors and consequently a much better understanding of the biology and molecular biology of ...

Lentiviral vectors are under intense scrutiny as unique candidate viral vector vaccines against tumor and aggressive pathogens because of their ability to initiate potent and durable specific immune responses. Strategies that alleviate safety concerns will facilitate the clinical developments involving lentiviral ...

... have been shown to facilitate vectoral transport and polarized diffusion. These include glycosylation, addition of lipid anchors, ... 16]. Notwithstanding these functional implications, the mechanisms reg...

Contents: Retrometabolic Approaches to Drug Targeting; Vector-Mediated Delivery of Opioid Peptides to the Brain; Conformationally Constrained Peptide Drugs Targeted at the Blood-Brain Barrier; Passive and Facilitative Transport of Nucleobases, Nucleosides...

... surface in multiple directions to form a active space around the water bowl inside the large cage facilitating free orientation of mosquitoes. Thus, the differential preference or ... ...

... individual founders and 50 with clip cages were set up for control and PEMV-infection treatments. Settling assays Because one ... ...

... plays important roles in facilitation of a bloodmeal, lubrication of mouthparts, and parasite transmission for some vector ... electrophoresisSaliva of hematophagous arthropods plays important roles in lu...

Ralstonia eutropha is a fascinating microorganism with a great scientific importance and an immense commercial potential. A new genetic transformation system for the organism would greatly facilitate the biological study and the molecular engineering of this organism. We have developed a versatile...

Although we strictly impose a polytropic relationship (with gamma .... A novel algorithm facilitates the wind vector analysis of the HARLIE data. ... from exchange of axial angular momentum between the solid mantle and the core. ...

The expression of foreign proteins in the fission yeast, Schizosaccharomyces pombe, is achieved by introducing an expression vector along with a transducing vector containing an autonomously replicating sequence. We created the expression vector pCMV-Leu2, carrying the LEU2 gene, which complements S. pombeleu1-32, and the transducing ...

Murine leukemia virus (MLV)-based retroviral vector is widely used for gene transfer. Efficient packaging of the genomic RNA is critical for production of high-titer virus. Here, we report that expression of the insulin-like growth factor II mRNA binding protein 1 (IMP1) enhanced the production of infectious MLV vector. Overexpression of IMP1 increased the ...

BACKGROUND: Although recombinant adenovirus vectors are attractive for use in gene expression studies and therapeutic applications, the construction of these vectors remains relatively time-consuming. We report here a strategy that simplifies the production of adenoviruses using the Cre-loxP system. MATERIALS AND METHODS: Full-length recombinant adenovirus ...

Lentiviral vectors are among the most efficient gene transfer tools for dividing and non-dividing cells. However, insertional mutagenesis has been observed in clinical trials with oncoretroviral vectors and this has prompted detailed study of genotoxicty of all integrating vectors. For many applications, avoiding integration is the ...

Understanding the mechanisms controlling vector-transmission of plant viruses requires integrating information from at least three different viewpoints: virus-vector interactions, plant-vector interactions and virus-plant interactions. While some of these aspects have been covered by past and present investigations, others have been ...

Biomaterials can potentially enhance the delivery of viral and nonviral vectors for both basic science and clinical applications. Vectors typically consist of nucleic acids (DNA, RNA) packaged with proteins, lipids, or cationic polymers, which facilitate cellular internalization and trafficking. These vectors can ...

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...

Several vectors that facilitate stable fluorescent labeling of Burkholderia pseudomallei and Burkholderia thailandensis were constructed. These vectors combined the effectiveness of the mini-Tn7 site-specific transposition system with fluorescent proteins optimized for Burkholderia spp., enabling bacterial tracking during cellular ...

nonviral vector systems have been used to facilitate stable gene delivery of the fumarylacetoacetate hydrolase gene in a mouse model of hereditary tyrosine- mia type 1 [9] and to restore permanently expression less active than CMV) comprised promoter sequences derived from Moloney murine leukemia virus, murine

BackgroundThe underlying goal of synthetic biology is to make the process of engineering biological systems easier. Recent work has focused on defining and developing standard biological parts. The technical standard that has gained the most traction in the synthetic biology community is the BioBrick standard for physical composition of genetic parts. Parts that conform to the BioBrick assembly ...

This brief technical note describes how to construct item vector plots for dichotomously scored items fitting the multidimensional three-parameter logistic model (M3PLM). As multidimensional item response theory (MIRT) shows promise of being a very useful framework in the test development life cycle, graphical tools that facilitate understanding of the ...

Acceleration technologies, in particular GPUs and Cell, are receiving considerable attention in modern-day HPC. Compared to classic accelerators and traditional CPUs, these devices not only exhibit higher compute density, but also sport significant memory bandwidth and vector-like capabilities to stream data at bandwidth of 100 GB/s or more. The latter qualifies such ...

The double-joint polymerase chain reaction (DJ-PCR) is a technique that can be used to construct vectors for targeted genome integration without laborious subcloning steps. Here we report the availability of plasmids that facilitate DJ-PCR-based construction of Neurospora crassa tagging vectors. These plasmids allow the creation of ...

We have developed a novel expression vector based on the bacteriophage phi 105, and employed it for the production of mutant beta-lactamases in Bacillus subtilis. Expression of the beta-lactamase-encoding gene was low when cloned into the prophage under the control of its own promoter. However, expression was considerably elevated when the gene was inserted into the phage ...

It has been previously shown that integrase-defective HIV-1-based gene vectors can serve, with moderate efficiency, as substrate for DNA transposition by a transiently expressed Sleeping Beauty (SB) transposase. Here, we describe the enhanced gene transfer properties of a HIV-1/SB hybrid vector that allows efficient DNA transposition, ...

Adenovirus (Ad) vectors have been developed as human immunodeficiency-1 (HIV-1) vaccine vectors because they consistently induce immune responses in preclinical animal models and human trials. Strong promoters and codon-optimization are often used to enhance vaccine-induced HIV-1 gene expression and immunogenicity. However, if the transgene is inherently ...

Adeno-associated virus serotype 5 (AAV5) is under development for gene-therapy applications for the treatment of cystic fibrosis. To elucidate the structural features of AAV5 that control its enhanced transduction of the apical surface of airway epithelia compared with other AAV serotypes, X-ray crystallographic studies of the viral capsid have been ...

The development of novel transformation vectors is essential to the improvement of plant transformation technologies. Here, we report the construction and testing of a new multifunctional dual binary vector system, pCLEAN, for Agrobacterium-mediated plant transformation. The pCLEAN vectors are based on the widely used pGreen/pSoup ...

The use of a nonviral magnetic vector, comprised of magnetic iron oxide nanoparticles (MNP), polyethylenimine (PEI), and plasmid DNA, for transfection of BHK21 cells under a magnetic field is presented. Four different vector configurations were studied by systematically varying the mixing order of MNP, PEI, and DNA. The assembly of the ...

We previously used directed evolution in human airway epithelia to create adeno-associated virus 2.5T (AAV2.5T), a highly infectious chimera of AAV2 and AAV5 with one point mutation (A581T). We hypothesized that the mechanism for its increased infection may be a higher binding affinity to the surface of airway epithelia than its parent AAV5. Here, we show ...

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Nonindigenous vectors that arrive, establish, and spread in new areas have fomented throughout recorded history epidemics of human diseases such as malaria, yellow fever, typhus, and plague. Although some vagile vectors, such as adults of black flies, biting midges, and tsetse flies, have dispersed into new habitats by flight or wind, human-aided transport ...

The interplay between vector and pathogen is essential for vector-borne disease transmission. Dissecting the molecular basis of refractoriness of some vectors may pave the way to novel disease control mechanisms. A pathogen often needs to overcome several physical barriers, such as the peritrophic matrix, midgut epithelium and salivary ...

We simultaneously transduced cells with three lentiviral gene ontology (LeGO) vectors encoding red, green or blue fluorescent proteins. Individual cells were thereby marked by different combinations of inserted vectors, resulting in the generation of numerous mixed colors, a principle we named red-green-blue (RGB) marking. We show that lentiviral ...

To facilitate a more efficient radiation and chemotherapy of mammary tumours, synthetic enhancer elements responsive to hypoxia and ionizing radiation were coupled to the mammary-specific minimal promoter of the murine whey acidic protein (WAP) encoding gene. The modified WAP promoter was introduced into a retroviral promoter conversion (ProCon) vector. ...

We have developed a DIY (Do It Yourself) series of genetic cassettes, which facilitate construction of novel versatile vectors for Alphaproteobacteria. All the cassettes are based on defined genetic modules derived from three natural plasmids of Paracoccus aminophilus JCM 7686. We have constructed over 50 DIY cassettes, which differ in structure and ...

RNA interference is the most rapid method for generation of conditional knockdown mutants in Trypanosoma brucei. The dual T7 promoter (pZJM) and the stem-loop vectors have been widely used to generate stable inducible RNAi cell lines with the latter providing tighter regulatory control. However, the steps for cloning stem-loop constructs are cumbersome requiring either ...

Abstract Viral vectors based on various naturally occurring adeno-associated virus (AAV) serotypes are among the most promising tools in human gene therapy. For the production of recombinant AAV (rAAV) vectors, researchers are focusing predominantly on cross-packaging an artificial AAV genome based on serotype 2 (AAV2) into capsids derived from other ...

Adenoviral vectors are widely used for cancer therapy and show a tumor-suppressing effect. However, bladder cancers are found to be resistant against infection of Ad5-derived adenoviral vector, limiting the application of the existing strategy of gene therapy. Therefore, efforts to develop novel types of adenoviral vector aimed for ...

BackgroundPlants are increasingly being examined as alternative recombinant protein expression systems. Recombinant protein expression levels in plants from Tobacco mosaic virus (TMV)-based vectors are much higher than those possible from plant promoters. However the common TMV expression vectors are costly, and at times technically challenging, to work ...

When processing Doppler optical coherence tomography images, there is a need to segment the Doppler signatures of the vessels. This can be used for visualization, for finding the center point of the flow areas or to facilitate the quantitative analysis of the vessel flow. We propose the use of a support-vector machine classifier in order to segment the ...

When processing Doppler optical coherence tomography images, there is a need to segment the Doppler signatures of the vessels. This can be used for visualization, for finding the center point of the flow areas or to facilitate the quantitative analysis of the vessel flow. We propose the use of a support-vector machine classifier in order to segment the ...

This paper discusses the compositional and phase relationships among minerals in which rare earth elements (REE) occur as essential constituents (e.g., bastnaesite, monazite, xenotime, aeschynite, allanite). Particular consideration is given to the vector representation of complex coupled substitutions in selected REE-bearing minerals and to the REE partitioning between ...

AbstractUnderstanding the structure and function of neural circuits is central is neuroscience research. To address the associated questions, new genetically encoded tools have been developed for mapping, monitoring, and manipulating neurons. Essential to implementation of these tools is their selective delivery to defined neuronal populations in the brain. This has been ...