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Genetics. 2006 Jun;173(2):769-77. Epub 2006 Mar 17.

Site-specific transformation of Drosophila via phiC31 integrase-mediated cassette exchange.

Author information

1
Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.

Abstract

Position effects can complicate transgene analyses. This is especially true when comparing transgenes that have inserted randomly into different genomic positions and are therefore subject to varying position effects. Here, we introduce a method for the precise targeting of transgenic constructs to predetermined genomic sites in Drosophila using the C31 integrase system in conjunction with recombinase-mediated cassette exchange (RMCE). We demonstrate the feasibility of this system using two donor cassettes, one carrying the yellow gene and the other carrying GFP. At all four genomic sites tested, we observed exchange of donor cassettes with an integrated target cassette carrying the mini-white gene. Furthermore, because RMCE-mediated integration of the donor cassette is necessarily accompanied by loss of the target cassette, we were able to identify integrants simply by the loss of mini-white eye color. Importantly, this feature of the technology will permit integration of unmarked constructs into Drosophila, even those lacking functional genes. Thus, C31 integrase-mediated RMCE should greatly facilitate transgene analysis as well as permit new experimental designs.

PMID:
16547094
PMCID:
PMC1526508
DOI:
10.1534/genetics.106.056945
[Indexed for MEDLINE]
Free PMC Article

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