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.
CITATION STYLE
Otabe, T., Nihongaki, Y., & Sato, M. (2023). A Split CRISPR–Cpf1 Platform for Inducible Gene Activation. In Methods in Molecular Biology (Vol. 2577, pp. 229–240). Humana Press Inc. https://doi.org/10.1007/978-1-0716-2724-2_16
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