Efficient Genome Modification by CRISPR-Cas9 Nickase With Minimal Off-Target Effects

Nat Methods. 2014 Apr;11(4):399-402. doi: 10.1038/nmeth.2857. Epub 2014 Mar 2.

Abstract

Bacterial RNA-directed Cas9 endonuclease is a versatile tool for site-specific genome modification in eukaryotes. Co-microinjection of mouse embryos with Cas9 mRNA and single guide RNAs induces on-target and off-target mutations that are transmissible to offspring. However, Cas9 nickase can be used to efficiently mutate genes without detectable damage at known off-target sites. This method is applicable for genome editing of any model organism and minimizes confounding problems of off-target mutations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • CRISPR-Cas Systems*
  • Deoxyribonuclease I / genetics
  • Deoxyribonuclease I / metabolism*
  • Embryo, Mammalian
  • Endonucleases / metabolism*
  • Gene Expression Regulation, Enzymologic
  • Genome, Bacterial*
  • HeLa Cells
  • Humans
  • Mice
  • Mutation
  • RNA, Bacterial

Substances

  • RNA, Bacterial
  • Endonucleases
  • Deoxyribonuclease I