Base editing with a Cpf1-cytidine deaminase fusion

Nat Biotechnol. 2018 Apr;36(4):324-327. doi: 10.1038/nbt.4102. Epub 2018 Mar 19.

Abstract

The targeting range of CRISPR-Cas9 base editors (BEs) is limited by their G/C-rich protospacer-adjacent motif (PAM) sequences. To overcome this limitation, we developed a CRISPR-Cpf1-based BE by fusing the rat cytosine deaminase APOBEC1 to a catalytically inactive version of Lachnospiraceae bacterium Cpf1. The base editor recognizes a T-rich PAM sequence and catalyzes C-to-T conversion in human cells, while inducing low levels of indels, non-C-to-T substitutions and off-target editing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • APOBEC-1 Deaminase / genetics*
  • Animals
  • CRISPR-Cas Systems / genetics
  • Clostridiales / enzymology
  • Deoxyribonucleases / genetics*
  • Gene Editing / methods*
  • Humans
  • Rats

Substances

  • Deoxyribonucleases
  • APOBEC-1 Deaminase
  • Apobec1 protein, rat