A split CRISPR-Cpf1 platform for inducible genome editing and gene activation

Nat Chem Biol. 2019 Sep;15(9):882-888. doi: 10.1038/s41589-019-0338-y. Epub 2019 Aug 12.

Abstract

The CRISPR-Cpf1 endonuclease has recently been demonstrated as a powerful tool to manipulate targeted gene sequences. Here, we performed an extensive screening of split Cpf1 fragments and identified a pair that, combined with inducible dimerization domains, enables chemical- and light-inducible genome editing in human cells. We also identified another split Cpf1 pair that is spontaneously activated. The newly generated amino and carboxyl termini of the spontaneously activated split Cpf1 can be repurposed as de novo fusion sites of artificial effector domains. Based on this finding, we generated an improved split dCpf1 activator, which has the potential to activate endogenous genes more efficiently than a previously established dCas9 activator. Finally, we showed that the split dCpf1 activator can efficiently activate target genes in mice. These results demonstrate that the present split Cpf1 provides an efficient and sophisticated genome manipulation in the fields of basic research and biotechnological applications.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anti-Bacterial Agents / pharmacology
  • Bacterial Proteins / metabolism*
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Endonucleases / metabolism*
  • Gene Editing
  • Gene Expression Regulation, Enzymologic / drug effects
  • HEK293 Cells
  • Humans
  • INDEL Mutation
  • Luciferases
  • Mice
  • Plasmids
  • RNA
  • Reproduction
  • Sirolimus / pharmacology

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • RNA
  • Luciferases
  • Cas12a protein
  • Endonucleases
  • Sirolimus