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

Wataru Fujii; Asuka Onuma; Koji Sugiura; Kunihiko Naito

doi:10.1016/j.bbrc.2014.01.141

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

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

Authors

Wataru Fujii¹, Asuka Onuma², Koji Sugiura², Kunihiko Naito³

Affiliations

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.

PMID: 24491566
DOI: 10.1016/j.bbrc.2014.01.141

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Base Sequence
Clustered Regularly Interspaced Short Palindromic Repeats*
Deoxyribonuclease I / genetics*
Gene Deletion
Gene Knock-In Techniques / methods*
Genome
Mice
Mice, Mutant Strains / genetics*
Molecular Sequence Data
RNA, Small Untranslated

Substances

Deoxyribonuclease I
RNA, Small Untranslated