Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects

Bin Shen, Wensheng Zhang, Jun Zhang, Jiankui Zhou, Jianying Wang, Li Chen, Lu Wang, Alex Hodgkins, Vivek Iyer, Xingxu Huang, William C. Skarnes

Research output: Contribution to journalArticlepeer-review

672 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)399-402
Number of pages4
JournalNature Methods
Volume11
Issue number4
DOIs
StatePublished - Apr 2014
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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