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- Title
Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects.
- Authors
Shen, Bin; Zhang, Wensheng; Zhang, Jun; Zhou, Jiankui; Wang, Jianying; Chen, Li; Wang, Lu; Hodgkins, Alex; Iyer, Vivek; Huang, Xingxu; Skarnes, William C
- 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.
- Subjects
EMBRYOS; BACTERIAL RNA; BACTERIAL genomes; MESSENGER RNA; ENDONUCLEASES; EUKARYOTES; MICROINJECTION (Cytology); LABORATORY mice; GENETIC mutation
- Publication
Nature Methods, 2014, Vol 11, Issue 4, p399
- ISSN
1548-7091
- Publication type
Article
- DOI
10.1038/nmeth.2857