A Split CRISPR-Cpf1 Platform for Inducible Gene Activation.

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.© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Citation

Takahiro Otabe, Yuta Nihongaki, Moritoshi Sato. A Split CRISPR-Cpf1 Platform for Inducible Gene Activation. Methods in molecular biology (Clifton, N.J.). 2023;2577:229-240

PMID: 36173577

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