Proceedings of the National Academy of Sciences of the United States of America, 24 August 2010, Vol.107(34), pp.15022-6
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.
Endonucleases -- Metabolism ; Gene Targeting -- Methods ; Zygote -- Metabolism
View this record in MEDLINE/PubMed