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

• DOI: 10.1038/srep11315
• Publication Date: 2015-06-11T14:13:11Z
• Authors: Masakazu Hashimoto; Tatsuya Takemoto
• Keywords: Mice, Inbred ICR; 0303 health sciences; Genome; Zygote; Article; Mice; 03 medical and health sciences; Electroporation; Animals; Female; Gene Knock-In Techniques; RNA, Messenger; CRISPR-Cas Systems

AbstractRecent 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.