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doi:10.1534/genetics.106.056945
A more recent version of this article appeared on June 1, 2006.
REGULAR RESEARCH PAPERS |
Site-specific transformation of Drosophila via 
C31 integrase-mediated cassette exchange
Jack R. Bateman 1, Anne M. Lee 1 and C.-Ting Wu 1*
1 Harvard Medical School
* To whom correspondence should be addressed. E-mail: twu{at}genetics.med.harvard.edu.
Submitted on February 8, 2006
Revised on March 9, 2006
Accepted on 14 March 2006
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.
Key Words: Drosophila, RMCE, phiC31 integrase, targeted transgenesis
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