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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.
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These DSBs can lead to highly specific gene edits when combined with HDR, or originate precise deletions in critical alleles through NHEJ.
“which can lead to highly specific gene edits when combined with HDR, or originate precise deletions in critical alleles through NHEJ [128,129].”
From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [129]
Pairing of Cas9n systems results in DSBs with gRNA-defined overhangs.
“The pairing of Cas9n systems results in DSBs with gRNA-defined overhangs”
From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [129]
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Metadata
- Authors
- Shen, B., Zhang, W., Zhang, J., Zhou, J., Wang, J., Chen, L., Wang, L., Hodgkins, A., Iyer, V., Huang, X., & Skarnes, W. C.
- Venue
- Nature Methods
- Published
- 2014-03-02
- Volume
- 11
- Issue
- 4
- Pages
- 399-402
- ISSN
- 1548-7091
- 1548-7105
- Cited by
- 769
- References
- 15
- Language
- en
- Licence
- www.springer.com/tdm (tdm)
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