TY - JOUR
T1 - Gene targeting by homologous recombination in mouse zygotes mediated by zinc-finger nucleases
AU - Meyer, Melanie
AU - De Angelis, Martin Hrabé
AU - Wurst, Wolfgang
AU - Kühn, Ralf
PY - 2010/8/24
Y1 - 2010/8/24
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=77957005768&partnerID=8YFLogxK
U2 - 10.1073/pnas.1009424107
DO - 10.1073/pnas.1009424107
M3 - Article
C2 - 20686113
AN - SCOPUS:77957005768
SN - 0027-8424
VL - 107
SP - 15022
EP - 15026
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 34
ER -