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Exploiting Position Effects And The Gypsy Retrovirus Insulator To Engineer Precisely Expressed Transgenes, Michele Markstein, Chrysoula Pitsouli, Christians Villalta, Susan E. Celniker, Norbert Perrimon
Exploiting Position Effects And The Gypsy Retrovirus Insulator To Engineer Precisely Expressed Transgenes, Michele Markstein, Chrysoula Pitsouli, Christians Villalta, Susan E. Celniker, Norbert Perrimon
Michele Markstein
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 …