transgenes

..We generated a collection of lines with precisely mapped attP sites that allow the insertion of transgenes into many different predetermined intergenic locations throughout the fly genome...

..strategies involve retroviral, lentiviral, adenoviral and plasmid transfection to deliver reprogramming factor transgenes. Although the latter two methods are transient and minimize the potential for insertion mutagenesis, they are ..

..This transgenesis platform should greatly facilitate structure/function analyses of most Drosophila genes...

..As an example, we show that the short stop gene is required in mushroom body neurons for the extension and guidance of their axons...

A major obstacle to creating precisely expressed transgenes lies in the epigenetic effects of the host chromatin that surrounds them...

..We have also generated transgenic rats that express GFP at high levels, suggesting that this technique can be used to produce other transgenic animal species...

At present, transgenes in Caenorhabditis elegans are generated by injecting DNA into the germline. The DNA assembles into a semistable extrachromosomal array composed of many copies of injected DNA...

..Upon differentiation, autosomal transgenes did not effect counting, but transgenic Xist RNA induced late replication and histone H4 hypoacetylation...

..In Brainbow transgenes, Cre/lox recombination is used to create a stochastic choice of expression between three or more fluorescent ..

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..SAHA dramatically improved the motor impairment in R6/2 mice, clearly validating the pursuit of this class of compounds as HD therapeutics...

..These results demonstrate that reprogramming human somatic cells does not require genomic integration or the continued presence of exogenous reprogramming factors and removes one obstacle to the clinical application of human iPS cells...

..Notably, the vectors did not detectably perturb endogenous miRNA expression or regulation of natural targets. The properties of miRNA-regulated vectors should allow for safer and more effective therapeutic applications...

..The resulting transgenes show robust expression in both germline and soma...

..Moreover it provides a platform to dissect the pathways of neurodegeneration in Alzheimer's disease and to develop novel therapeutic interventions...

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..The safety of the intervention and the stability of the improvement in visual and retinal function in these subjects support the use of AAV-mediated gene augmentation therapy for treatment of inherited retinal diseases...

The residual presence of integrated transgenes following the derivation of induced pluripotent stem (iPS) cells is highly undesirable...

..We show that reporter gene expression was responsive to doxycycline and that the resulting transgenic human ES cells retain their self-renewal capacity and pluripotency...

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..In the future, this technology will allow effective comparisons and informed choices to be made, potentially leading to complete transmission blockade...

..Adenoviral reprogramming may provide an improved method for generating and studying patient-specific stem cells and for comparing embryonic stem cells and iPS cells...

..Ability to transfer foreign genes into human ES cells has potential relevance for the development of gene and cell-based therapies...

The introduction of two transgenes into one animal is increasingly common as transgenic experiments become more sophisticated. In this study we examine two strategies for creating double transgenic founders from a single microinjection...

..These studies extend the understanding of the pluripotent genetic regulatory network within which the Sox2-Oct4 complex are at the top of the regulatory hierarchy...

..25 and 10.5. These data suggest that arterial pole myocardium originates outside the classical heart field...

Here we describe the sustained expression of transgenes introduced into human embryonic stem (ES) cells using self-inactivating lentiviral vectors...

..However, the conventional electroporation technique fails to retain the expression of transgenes for more than several days because transgenes are not integrated into the genome...

..remove this roadblock, a macrophage-specific marker was identified (mpeg1) and its promoter used in mpeg1-driven transgenes. mpeg1-driven transgenes are expressed in macrophage-lineage cells that do not express neutrophil-marking ..

..Thus, trichromacy can arise from a single addition of a third cone class and it does not require an early developmental process. This provides a positive outlook for the potential of gene therapy to cure adult vision disorders...

..The increased strength and reliability of these optimized reagents overcome many of the previous limitations of these methods and will facilitate genetic manipulations of greater complexity and sophistication...

Using a two-component transgene system involving two epiallelic variants of the invertedly repeated transgenes in locus 1 (Lo1) and a homologous single-copy transgene locus 2 (Lo2), we have studied the stability of the methylation ..

..This qPCR methodology offers specific, objective, and rapid quantitation, uncomplicated by tissue autofluorescence, and should be readily transferable to other in vivo models to quantitate the biolocalization of transplanted cells...

..Our data suggest that minicircles are capable of expressing high and persistent levels of therapeutic products in vivo and have a great potential to serve as episomal vectors for the treatment of a wide variety of diseases...

..HEN1 contribute to resistance against cucumber mosaic virus, but not to siRNA and IR-PTGS triggered by hairpin transgenes directly producing perfect dsRNA; and iv) the actions of HEN1 in miRNA/development and siRNA/S-PTGS can be ..

..b>Transgenes may persist to and spread in wild populations after gene flow, resulting unwanted ecological consequences...

..The artificial methylation was reprogrammed in the early embryo, leading to founders that express the transgenes. We also noted differences in transgene insertion number and structure (single-insert versus concatemer) based ..

..By broadening the antigenic peptide repertoire, nardilysin and TOP strengthen the immune defense against intracellular pathogens and cancer...

..Mutations were mapped and genes cloned by both traditional and whole genome sequencing approaches. The methodologies and the mutant alleles discovered will be instrumental in further studies of de novo DNA methylation...

..In addition, transgenes integrated as repeated arrays are susceptible to repeat-induced gene silencing...

..The purpose of this study was to test whether delivery of functional GC1 to cone cells of the postnatal GC1KO mouse could restore function to these cells...

..The genetic toolbox for Tribolium research has expanded in the past years but spatio-temporally controlled misexpression of genes has not been possible so far...

..RNAi-mediated suppression of EZH led to defects in the silencing of transgenes and retrotransposons as well as to a global increase in histone post-translational modifications associated with ..

..However, regardless of the mode of delivery, concerns continue to be raised by the presence of vector sequences detected at distant sites...

The schistosome egg represents an attractive developmental stage at which to target transgenes because of the high ratio of germ to somatic cells, because the transgene might be propagated and amplified by infecting snails with the ..

..This demonstrates that an RMCE system in the context of a viral vector allows the site directed insertion of a transgene into a defined genomic site...

..Specific inhibition of gene expression by RNAi has been shown to be suitable for a multitude of phytopathogenic filamentous fungi. However, double-stranded (ds)RNA/small interfering (si)RNA silencing effect has not been observed in vivo...

..This work smoothes the way for a novel method of misexpression of gene products in zebrafish in order to analyze the function of genes in developmental processes...

..Exemplified by Chinese hamster ovary cells, the strategy proves to be successful even for cell lines with an unstable genotype...

..We conclude that myosin VI does not need to dimerize via the predicted coiled coil to stabilize actin in vivo...

..Furthermore, these results indicate that gene delivery to the outer nuclear layer may be achieved by noninvasive intravitreal AAV administration in the diseased state...

..Importantly, in mammals, four FOXO genes have overlapping and different functions, and in C. elegans, a single FOXO/DAF-16 uses distinct isoforms to fine-tune the IIS-mediated processes in the context of a whole organism...

..These properties are achieved in the absence of classic enhancer activity and therefore may confer a high safety profile...

..Such an increase in the bioavailability of adenovirus, coupled with substantial decreases in toxicity and unwanted transgene expression is an important step towards producing systemically available tumour-targeted viruses...

..Cautions should be taken to choose the optimal promoter when lentiviruses are used for genetic modification of human ES cells...

..Here we describe the GAL4 system highlighting the properties that make it a powerful tool for the analysis of gene function in Drosophila and higher organisms...

..Together, these results further established that the DNA silencing effect was mediated by a covalent linkage of the expression cassette and the bacteria DNA elements...

..These results identify a common mechanism for sexual and species recognition regulated by cuticular hydrocarbons...

..Our results hold promise for future clinical trials of cone-directed gene therapy in achromatopsia and other cone-specific disorders...

..CyMATE is freely available for non-commercial use at http://www.gmi.oeaw.ac.at/CyMATE...

Inserting transgenes into bacterial chromosomes is generally quite involved, requiring a selection for cells carrying the insertion, usually for drug-resistance, or multiple cumbersome manipulations, or both...

..This amplification mechanism substantially augments the potency of RNAi-based surveillance, while ensuring that the RNAi machinery will focus on expressed mRNAs...

..The large size of the BAC transgenes ensures the presence of most, if not all, regulatory elements and results in expression that closely matches ..

..Hence, the setting of the Hox anterior expression border is not a simple function of the affinity of retinoid receptors for their cognate element...

..Further experiments address potential mechanisms of cooperativity between the two transgenes. In these bitransgenic mice, we have recapitulated two common genetic lesions that occur in human breast cancer ..

..Therefore, these elements may be very useful tools in gene therapy applications for ensuring high-level and stable expression of transgenes.

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..The vector production and titration protocol described here can be completed within 8 d...

..This identification of a differentiated pancreatic cell type as an in vivo progenitor of all differentiated pancreatic cell types has implications for a potential expandable source for new islets for replenishment therapy for diabetes...

..The most commonly used techniques for producing transgenic mice involves either the pronuclear injection of transgenes into fertilized oocytes or embryonic stem cell-mediated gene targeting...

..Here we describe an efficient method for the modification of previously inserted transgenes by the use of the site-specific integration system from phage phiC31 in a tephritid pest species, the ..

..Plants expressing human Bcl-2 and Bcl-xl, nematode CED-9, or baculovirus Op-IAP transgenes conferred heritable resistance to several necrotrophic fungal pathogens, suggesting that disease development ..

..The TARGET system offers general utility in simultaneously addressing issues of when and where gene products are required...

..Although YAC and cosmid Xist-containing transgenes have previously been reported to be capable of cis inactivation and counting, the transgenes were all present as ..

..A chondrogenic stimulus, e.g. by transfer of BMP-2 cDNA, appears to be required for initiation and support of chondrogenic differentiation...

..Collectively, an improvement in nuclear decondensation and the diminution of the interaction between vector and mRNA is essential for the development of new generations of non-viral vectors...

..chromosomes in host cells and should be free from potential insertional mutagenesis problems of conventional transgenes. Therefore, HACs have been proposed as alternative implements to cell-mediated gene therapy...

..Marker-free homozygous progeny plants were efficiently obtained, regardless of which promoter was used. In addition, simplification of complex transgene loci was observed...

..Concluding remarks address some of the successes in preclinical animals, and the recent transition of lentiviral vectors to human clinical trials as therapy for a variety of infectious and genetic diseases...

..In this review these newly developed strategies are discussed...

..of the distinct members of this family, GUS reporter fusion genes were constructed, and plants expressing these transgenes were characterized to reveal spatial and temporal patterns of expression...

We generated double-transgenic mice carrying cointegrated tissue-specific Gal4 and Gal4 reporter transgenes to direct transgene overexpression in the mononuclear phagocyte system (MPS)...

..Our results also highlight the utility of focused screens in the mouse, in addition to the large-scale and broadly targeted screens that are being carried out at mutagenesis centers...

..Germline specific promoters are an essential component of potential vector control strategies which function by genetic drive, however suitable promoters are not currently available for the main human malaria vector Anopheles gambiae...

..We analyze the dispersal of maize transgenes via seeds in Mexico, the crop's cradle.

..These findings support a new dual model for Myc chromatin function with important implications for the role of Myc in cancer and stem cell biology, including that of induced pluripotent stem cells...

..In conclusion, nonviral gene transfer can produce a functional cardiac biological pacemaker regulated by sympathetic input, which improves life expectancy in a mouse model of CAVB...

..Using this improved approach, we also generated hES-like cells from homozygous fibroblasts containing the sickle cell anemia mutation Hemoglobin Sickle. Disclosure of potential conflicts of interest is found at the end of this article...

..any desired gene product can be generated by crossing MAZe fish with other lines containing UAS-driven transgenes. The number of clones induced, and their time of origin, could be varied by adjusting heat-shock timing and ..

..In this organism, RNAi can be triggered either by high-copy, truncated transgenes or by directly feeding cells with double-stranded RNA (dsRNA)...

..This selection system allows hands-off maintenance and enrichment of transgenic worms carrying non-integrated transgenes on selective plates...

..Our findings promise to facilitate the development of miRNA-regulated rAAV for CNS-targeted gene delivery and other applications...

..Therefore, the combination of using mGS cells with HACs provides a new tool for gene and cell therapies. The next step is to demonstrate functional restoration using animal models for future gene therapy...

..However, type I IFN might modulate antitumor and antiadenoviral immune responses and thus influence the outcome of gene immunotherapy...

..In addition, transgene expression was observed for at least 6 months. This HF stem cell-targeted gene transfer and reconstitution system provides reliable gene-function analysis and gene therapy...

..This is the first study to document noninvasive monitoring of long-term transgene expression in the adult mouse brain and provides the basis for applying BLI in the study of brain disease and gene therapeutic strategies...

..Overall, compared to Ad5 vectors, Ad5/35 vectors had a better safety profile, reflected by lower serum levels of proinflammatory cytokines...

..Adenoviral vectors containing these enhanced promoters may be useful in glioma gene therapy...

Recombinant adeno-associated virus vectors based on serotype 2 (rAAV2) have been used to deliver transgenes to the airways in a variety of pre-clinical and clinical studies...

..CONCLUSIONS: The MESV-based retroviral systems, when combined with hormone inducible Cre, represent efficient tools for the transfer of Cre activity in ES cells...

..of this award, we have characterized mouse models of lung adenocarcinoma initiated by doxycycline-inducible transgenes encoding the two common forms of mutant EGFR found in the corresponding type of human cancer...

..These transgenes were subsequently shown to confer susceptibility to collagen induced arthritis in normally resistant B10.M...

..In fungi, plants, and animals the introduction of transgenes can lead to the silencing of homologous sequences by a variety of epigenetic phenomena operating at both ..

..These elements will be defined by studying the expression of Collal CAT reporter transgenes bearing various segments of the 5' promoter region as well as specific deletions of the first intron...

..Specific Aim 1 is to deliver transgenes to rhesus monkey preimplantation embryos with Epstein-Barr Virus (EBV)-based and retrovirus-based vectors...

..assess the role of TRAF proteins in CD40 mediated isotype switching, we have examined the ability of mutant CD40 transgenes that lack one or both TRAF binding sites (ATRAF6, ATRAF2/3 and ATRAFs respectively) to reconstitute isotype ..

..assess the role of TRAF proteins in CD40 mediated isotype switching, we have examined the ability of mutant CD40 transgenes that lack one or both TRAF binding sites (deltaTRAF6, deltaTRAF2/3 and deltaTRAFs respectively) to reconstitute ..

..To better understand the role of B lymphocytes in IDDM, NOD mice were produced that express immunoglobulin transgenes (Tg) from anti-insulin mab125...

..Our long term goal is to combine the insulin 1 and 2 knockouts with transgenes encoding altered insulin B:9-23 sequences in the same NOD strain and evaluate whether the B:9-23 native sequence ..

..The anti-HIV vectors we propose to test contain MGMT transgenes to allow in vivo selection post-transplantation...

..The anti-HIV vectors we propose to test contain MGMT transgenes to allow in vivo selection post-transplantation...

..and validation of the Sleeping Beauty (SB) transposon system for its efficacy to catalyze integration of transgenes to cure mucopolysaccharidosis diseases in murine and canine model systems...

..However, current iPSCs suffer from major drawbacks including multiple viral integrations and remaining transgenes at various chromosomal locations, any of which may cause unpredictable genetic dysfunction and/or tumor ..

..While these systems have been proven effective, the requirements prohibit differential regulation of multiple transgenes. Additionally, the requirement of co-delivering a trans-activator adds significantly to the payload required of ..

..The DMD is essential to imprint H19 transgenes as well as the endogenous H19 gene, and the linked and oppositely imprinted Igf2 gene...

..We have developed lines of Xenopus laevis in which transgenes both cause rod photoreceptors to degenerate in a manner that resemble human retinitis pigmentosa as well as ..

..For A, transgenes with cell type-specific promoter/enhancers will be used to drive LeprB expression...

..For Aims 2-5, mice will be created in which transgenes can be inducibly expressed in a lineage-specific manner...

..The next challenge to implementation of transgenic mosquitoes is development of tools to drive the spread of transgenes from small numbers of laboratory- manipulated mosquitoes into field populations...

..b>Transgenes must be actively driven into natural Anopheles populations to high frequency for transgenic strategies to be ..

..Using a number of transgenes designed to alter IGF expression, we will study in utero actions and mechanisms of action of IGFs in cultured ..