Efficient generation of genome-modified mice via offset-nicking by CRISPR/Cas system.
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.
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.
Wataru
W
Fujii
Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Tokyo 113-8657, Japan.
Asuka
A
Onuma
Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Tokyo 113-8657, Japan.
Koji
K
Sugiura
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.
Kunihiko
K
Naito
D000818:Animals; D001483:Base Sequence; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D003850:Deoxyribonuclease I / Q000235:genetics*; D017353:Gene Deletion; D055879:Gene Knock-In Techniques / Q000379:methods*; D016678:Genome; D051379:Mice; D008817:Mice, Mutant Strains / Q000235:genetics*; D008969:Molecular Sequence Data; D058727:RNA, Small Untranslated
D003850:Deoxyribonuclease I; D058727:RNA, Small Untranslated
CRISPR/Cas9; Genome-modified animals; Nickase; Off-target