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Generation of mutant mice with minimal off-target effects by microinjection of one-cell embryos with paired sgRNAs and Cas9 nickase

External protocol Created on 30 Apr 2014

Authors

William C. Skarnes & Xingxu Huang

Summary

Cas9 nickase and paired sgRNAs can be used to efficiently modify the genome of one-cell embryos with minimal off-target damage1. This protocol describes the design and construction of paired sgRNAs plasmids, preparation of sgRNAs and Cas9-D10A mRNAs for the injection of one-cell embryos, and, genotyping of founder animals.

Further details

The protocol was published on Protocol Exchange on 3 March 2014. To see the entire protocol, click on the source link.

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