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.
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The datasets generated during this study are available from the corresponding author upon request.
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This work was supported by START and CREST grants (no. JPMJCR1653) from Japan Science and Technology Agency (JST) and grants for Project for Cancer Research and Therapeutic Evolution (no. 17cm0106415h0002) from Japan Agency for Medical Research and Development (AMED) to M.S. This work was also supported by grants from Japan Society for the Promotion of Science (JSPS) to M.S. and JSPS Research Fellowships for Young Scientists to Y.N. (no. 15J05897).
The authors declare no competing interests.
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