Accelerated homologous recombination and subsequent genome modification in Drosophila.

ABSTRACT: Gene targeting by 'ends-out' homologous recombination enables the deletion of genomic sequences and concurrent introduction of exogenous DNA with base-pair precision without sequence constraint. In Drosophila, this powerful technique has remained laborious and hence seldom implemented. We describe a targeting vector and protocols that achieve this at high frequency and with very few false positives in Drosophila, either with a two-generation crossing scheme or by direct injection in embryos. The frequency of injection-mediated gene targeting can be further increased with CRISPR-induced double-strand breaks within the region to be deleted, thus making homologous recombination almost as easy as conventional transgenesis. Our targeting vector replaces genomic sequences with a multifunctional

SUBMITTER: Baena-Lopez LA 

PROVIDER: S-EPMC3833436 | biostudies-literature | 2013 Dec

REPOSITORIES: biostudies-literature