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

doi:10.1038/nmeth.2857

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 journal › Article › peer-review

724 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 language	English (US)
Pages (from-to)	399-402
Number of pages	4
Journal	Nature Methods
Volume	11
Issue number	4
DOIs	https://doi.org/10.1038/nmeth.2857
State	Published - Apr 2014
Externally published	Yes

ASJC Scopus subject areas

Biotechnology
Biochemistry
Molecular Biology
Cell Biology

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10.1038/nmeth.2857

Cite this

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title = "Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects",

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.",

author = "Bin Shen and Wensheng Zhang and Jun Zhang and Jiankui Zhou and Jianying Wang and Li Chen and Lu Wang and Alex Hodgkins and Vivek Iyer and Xingxu Huang and Skarnes, \{William C.\}",

note = "Funding Information: We thank members of the entire Huang laboratory for their support and advice, G. Tischler for the C++ alignment code, and J. Liu and C. Gao for help with deep sequencing. This work was supported by grants from the 973 program (2010CB945101 and 2011CB944301) and a core grant from the Wellcome Trust.",

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AU - Shen, Bin

AU - Zhang, Wensheng

AU - Zhang, Jun

AU - Zhou, Jiankui

AU - Wang, Jianying

AU - Chen, Li

AU - Wang, Lu

AU - Hodgkins, Alex

AU - Iyer, Vivek

AU - Huang, Xingxu

AU - Skarnes, William C.

N1 - Funding Information: We thank members of the entire Huang laboratory for their support and advice, G. Tischler for the C++ alignment code, and J. Liu and C. Gao for help with deep sequencing. This work was supported by grants from the 973 program (2010CB945101 and 2011CB944301) and a core grant from the Wellcome Trust.

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N2 - 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.

AB - 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.

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

Abstract

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