Gene targeting by homologous recombination in mouse zygotes mediated by zinc-finger nucleases.

ABSTRACT: Gene targeting by homologous recombination in embryonic stem cells is extensively used to generate specific mouse mutants. However, most mammalian species lack tools for targeted gene manipulation. Since double-strand breaks strongly increase the rate of homologous recombination at genomic loci, we explored whether gene targeting can be directly performed in zygotes by the use of zinc-finger nucleases. Here we report that gene targeting is achieved in 1.7-4.5% of murine one-cell embryos upon the coinjection of targeting vectors with zinc-finger nucleases, without preselection. These findings enable the manipulation of the mammalian germ line in a single step in zygotes, independent of ES cells.

SUBMITTER: Meyer M 

PROVIDER: S-EPMC2930558 | biostudies-other | 2010 Aug

REPOSITORIES: biostudies-other