Electroporation Enables the Efficient mRNA Delivery Into the Mouse Zygotes and Facilitates CRISPR/Cas9-based Genome Editing

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems*
  • Electroporation*
  • Female
  • Gene Knock-In Techniques / methods*
  • Genome*
  • Mice
  • Mice, Inbred ICR
  • RNA, Messenger / chemistry*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Zygote*

Substances

  • RNA, Messenger