A Split CRISPR–Cpf1 Platform for Inducible Gene Activation

Takahiro Otabe, Yuta Nihongaki, Moritoshi Sato

Research output: Chapter in Book/Report/Conference proceedingChapter


The CRISPR–Cpf1 also known as Cas12a is an RNA-guided endonuclease similar to CRISPR–Cas9. Combining the CRISPR–Cpf1 with optogenetics technology, we have engineered photoactivatable Cpf1 (paCpf1) to precisely control the genome sequence in a spatiotemporal manner. We also identified spontaneously activated split Cpf1 and thereby developed a potent dCpf1 split activator, which has the potential to activate endogenous target genes. Here we describe a method for optogenetic endogenous genome editing using paCpf1 in mammalian cells. Furthermore, we show a method for endogenous gene activation using dCpf1 split activator in mammalian cells and mice.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages12
StatePublished - 2023

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • CRISPR–Cpf1 (also known as Cas12a)
  • crRNA
  • Gene activation
  • Genome editing
  • Genomic PCR
  • Hydrodynamic tail vein injection
  • In vivo gene activation
  • NHEJ
  • Optogenetics
  • T7E1

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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