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

Shen, Bin; Zhang, Wensheng; Zhang, Jun; Zhou, Jiankui; Wang, Jianying; Chen, Li; Wang, Lu; Hodgkins, Alex et al. · Nat Methods · 2014

basic_science · Level V

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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.

Medical subject headings