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Markstein, M., Pitsouli, C., Villalta, C., Celniker, S.E., Perrimon, N. (2008). Exploiting position effects and the gypsy retrovirus insulator to engineer precisely expressed transgenes.  Nat. Genet. 40(4): 476--483.
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Publication Type
Research paper

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.

PubMed ID
PubMed Central ID
PMC2330261 (PMC) (EuropePMC)
Related Publication(s)

Delivery codes for fly transgenics.
Tubon and Yin, 2008, Nat. Genet. 40(4): 383--384 [FBrf0216072]

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    Language of Publication
    Additional Languages of Abstract
    Parent Publication
    Publication Type
    Nat. Genet.
    Nature Genetics
    Publication Year
    1061-4036 1546-1718
    Data From Reference
    Alleles (9)
    Balancers (1)
    Genes (38)
    Natural transposons (2)
    Insertions (51)
    Experimental Tools (2)
    Transgenic Constructs (9)