1. Exploiting position effects and the gypsy retrovirus insulator to engineer precisely expressed transgenes
- Author
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Markstein, M., Pitsouli, Chrysoula, Villalta, C., Celniker, S. E., Perrimon, N., and Pitsouli, Chrysoula [0000-0003-4074-9684]
- Subjects
insulator element ,Genome, Insect ,Genome ,Wing ,Animals, Genetically Modified ,Retrovirus ,Drosophila Proteins ,Wings, Animal ,Tissue Distribution ,Transgenes ,transcription factor GAL4 ,Recombination, Genetic ,Regulation of gene expression ,Genetics ,Receptors, Notch ,article ,luciferase ,Chromatin ,Drosophila melanogaster ,Phenotype ,Position effect ,priority journal ,Myogenic Regulatory Factors ,Larva ,Attachment Sites, Microbiological ,Drosophila ,Female ,Insulator Elements ,Plasmids ,Saccharomyces cerevisiae Proteins ,gene locus ,phenotype ,Transgene ,DNA transcription ,Molecular Sequence Data ,Biology ,Article ,Animals ,controlled study ,HSP70 Heat-Shock Proteins ,Epigenetics ,nonhuman ,transgene ,nucleotide sequence ,biology.organism_classification ,Cytoskeletal Proteins ,Retroviridae ,Gene Expression Regulation ,gene expression - Abstract
A major obstacle to creating precisely expressed transgenes lies in the epigenetic effects of the host chromatin that surrounds them. Here we present a strategy to overcome this problem, employing a Gal4-inducible luciferase assay to systematically quantify position effects of host chromatin and the ability of insulators to counteract these effects at phiC31 integration loci randomly distributed throughout the Drosophila genome. We identify loci that can be exploited to deliver precise doses of transgene expression to specific tissues. Moreover, we uncover a previously unrecognized property of the gypsy retrovirus insulator to boost gene expression to levels severalfold greater than at most or possibly all un-insulated loci, in every tissue tested. These findings provide the first opportunity to create a battery of transgenes that can be reliably expressed at high levels in virtually any tissue by integration at a single locus, and conversely, to engineer a controlled phenotypic allelic series by exploiting several loci. The generality of our approach makes it adaptable to other model systems to identify and modify loci for optimal transgene expression. © 2008 Nature Publishing Group. 40 476 483 Cited By :231
- Published
- 2008