A powerful method combining homologous recombination and site-specific recombination for targeted mutagenesis in Drosophila.

ABSTRACT: Gene targeting provides a powerful tool for dissecting gene function. However, repeated targeting of a single locus remains a practice mostly limited to unicellular organisms that afford simple targeting methodologies. We developed an efficient method to repeatedly target a single locus in Drosophila. In this method, which we term "site-specific integrase mediated repeated targeting" (SIRT), an attP attachment site for the phage phiC31 integrase is first targeted to the vicinity of the gene of interest by homologous recombination. All subsequent modifications of that gene are introduced by phiC31-mediated integration of plasmids carrying an attB attachment site and the desired mutation. This highly efficient integration results in a tandem duplication of the target locus, which is then red

SUBMITTER: Gao G 

PROVIDER: S-EPMC2529331 | biostudies-literature | 2008 Sep

REPOSITORIES: biostudies-literature