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

Melanie Meyer, Martin Hrabé De Angelis, Wolfgang Wurst, Ralf Kühn

Research output: Contribution to journalArticlepeer-review

246 Scopus citations

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.

Original languageEnglish
Pages (from-to)15022-15026
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume107
Issue number34
DOIs
StatePublished - 24 Aug 2010

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