Efficient Generation of Genome-Modified Mice Using Campylobacter jejuni-Derived CRISPR/Cas

International Journal of Molecular Sciences
Wataru FujiiKunihiko Naito

Abstract

Mammalian zygote-mediated genome-engineering by Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/Cas is currently used for the generation of genome-modified animals. Here, we report that a Campylobacter jejuni-derived orthologous CRISPR/Cas system recognizes a 5'-NNNVRYAC sequence as a protospacer-adjacent motif in mouse zygotes, and is applicable for efficient generation of knockout mice. Moreover, this novel CRISPR/Cas can be used for zygote-mediated knock-in at a unique locus, suggesting that this system could help to expand the feasibility of the zygote-mediated generation of genome-modified animals.

References

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Jun 24, 2017·Methods in Molecular Biology·Wataru Fujii

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Methods Mentioned

BETA
Transfection
genotyping
PCR

Software Mentioned

ssODN
IDT Codon Optimization Tool

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