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Sci Rep. 2015 Jun 11;5:11315. doi: 10.1038/srep11315.

Electroporation enables the efficient mRNA delivery into the mouse zygotes and facilitates CRISPR/Cas9-based genome editing.

Author information

1
Division of Developmental Biology, Graduate School of Medicine, Chiba University, Inohana 1-8-1, Chuo-ku, Chiba 260-8670, Japan.
2
Division of Embryology, Fujii Memorial Institute of Medical Sciences, Tokushima University, 3-18-15 kuramoto-cho, Tokushima 770-8503, Japan.

Abstract

Recent use of the CRISPR/Cas9 system has dramatically reduced the time required to produce mutant mice, but the involvement of a time-consuming microinjection step still hampers its application for high-throughput genetic analysis. Here we developed a simple, highly efficient, and large-scale genome editing method, in which the RNAs for the CRISPR/Cas9 system are electroporated into zygotes rather than microinjected. We used this method to perform single-stranded oligodeoxynucleotide (ssODN)-mediated knock-in in mouse embryos. This method facilitates large-scale genetic analysis in the mouse.

PMID:
26066060
PMCID:
PMC4463957
DOI:
10.1038/srep11315
[Indexed for MEDLINE]
Free PMC Article

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