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.

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, Guide / genetics*

Substances

  • RNA, Guide
  • Deoxyribonuclease I