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Biochem Biophys Res Commun. 2014 Mar 21;445(4):791-4. doi: 10.1016/j.bbrc.2014.01.141. Epub 2014 Jan 31.

Efficient generation of genome-modified mice via offset-nicking by CRISPR/Cas system.

Author information

1
Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Tokyo 113-8657, Japan. Electronic address: awtrfj@mail.ecc.u-tokyo.ac.jp.
2
Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Tokyo 113-8657, Japan.
3
Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Tokyo 113-8657, Japan. Electronic address: aknaito@mail.ecc.u-tokyo.ac.jp.

Abstract

The mammalian zygote-mediated CRISPR/Cas system can efficiently generate targeted genome-modified animals. However, this system is limited by the risk of off-target mutations. Here we show that offset-nicking by Cas9 nickase and paired gRNAs allows us to generate region deleted mice and targeted knock-in mice without off-target mutations.

KEYWORDS:

CRISPR/Cas9; Genome-modified animals; Nickase; Off-target

PMID:
24491566
DOI:
10.1016/j.bbrc.2014.01.141
[Indexed for MEDLINE]
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