An inducible CRISPR/Cpf1 system for multiplex gene activation

Abstract:

In order to develop highefficiency endogenous multigene activation tools, and based on the CRISPR/Cpf1 system, which can promote the self-cleavage and release of multiple crRNAs in pre-crRNA, the TRE3G-DD-dCpf1-VPR activation system was constructed. In this system, the sitedirected mutation of the nuclease region of As/Fn/LbCpf1 to form dCpf1, and The VPR with DD-domain the tetracycline-controlled operon were introduced at the C and N terminal, respectively. The results showed that the transcription level of Oct4 gene was increased by 104 times, and the transcription levels of Myod1 and Oct4 genes can achieve different levels under the control of Dox and TMP single drugs respectively. Under the condition of adding dual drugs, Myod1 and Oct4 genes can be increased by more than 80 and 120 times respectively. It proves that the system can induce endogenous multi-gene activation quickly and efficiently by targeting DNA.