title: Electroporation enables the efficient mRNA delivery into the mouse zygotes and facilitates CRISPR/Cas9-based genome editing.
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.
authors.affiliation: Division of Developmental Biology, Graduate School of Medicine, Chiba University, Inohana 1-8-1, Chuo-ku, Chiba 260-8670, Japan.
authors.forename: Masakazu
authors.initials: M
authors.lastname: Hashimoto
authors.affiliation: Division of Embryology, Fujii Memorial Institute of Medical Sciences, Tokushima University, 3-18-15 kuramoto-cho, Tokushima 770-8503, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Takemoto
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D018274:Electroporation; D005260:Female; D055879:Gene Knock-In Techniques / Q000379:methods*; D016678:Genome; D051379:Mice; D008813:Mice, Inbred ICR; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015053:Zygote
chemical_list: D012333:RNA, Messenger