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.

CRISPR-Cpf1 核酸内切酶最近已被证明是操纵靶向基因序列的强大工具。在这里，我们对分裂的 Cpf1 片段进行了广泛的筛选，并确定了一对与诱导型二聚化结构域相结合的片段，可以在人类细胞中进行化学和光诱导的基因组编辑。我们还发现了另一个自发激活的分裂 Cpf1 对。自发激活的分裂 Cpf1 新产生的氨基和羧基末端可以重新用作人工效应域的从头融合位点。基于这一发现，我们生成了一种改进的分裂 dCpf1 激活剂，它有可能比以前建立的 dCas9 激活剂更有效地激活内源基因。最后，我们证明了分裂的 dCpf1 激活剂可以有效地激活小鼠中的靶基因。