Highly Efficient Cpf1-Mediated Gene Targeting in Mice Following High Concentration Pronuclear Injection

G3 (Bethesda). 2017 Feb 9;7(2):719-722. doi: 10.1534/g3.116.038091.

Abstract

Cpf1 has emerged as an alternative to the Cas9 RNA-guided nuclease. Here we show that gene targeting rates in mice using Cpf1 can meet, or even surpass, Cas9 targeting rates (approaching 100% targeting), but require higher concentrations of mRNA and guide. We also demonstrate that coinjecting two guides with close targeting sites can result in synergistic genomic cutting, even if one of the guides has minimal cutting activity.

Keywords: CRISPR; Cpf1 nuclease; genome editing; mouse.

MeSH terms

  • Acidaminococcus / enzymology
  • Acidaminococcus / genetics
  • Animals
  • Bacterial Proteins / genetics*
  • CRISPR-Cas Systems / genetics
  • Endonucleases / genetics*
  • Gene Editing / methods*
  • Gene Targeting / methods*
  • Mice
  • RNA, Guide, CRISPR-Cas Systems / genetics*
  • RNA, Messenger / genetics

Substances

  • Bacterial Proteins
  • RNA, Guide, CRISPR-Cas Systems
  • RNA, Messenger
  • Cas12a protein
  • Endonucleases