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

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

Author information

1
1] Ministry of Education Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, Nanjing, China. [2].
2
1] Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK. [2].
3
Ministry of Education Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, Nanjing, China.
4
Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China.
5
Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.

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.

PMID:
24584192
DOI:
10.1038/nmeth.2857
[Indexed for MEDLINE]
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