A chemical CRISPR off switch efficiently controls gene editing
Xingyu Liu, Wei Xiong, Qian-Qian Qi, Huimin Ji, Yutong Zhang, Huajun Lei, Jian Liu, Ping Yin, Tian Tian, Xiang Zhou
Abstract
Chemical methods for regulating CRISPR activity hold great potential for spatiotemporal control of gene editing technologies. Here we report a small-molecule strategy to switch off CRISPR functionality. We develop clickable RNAs by modifying RNAs with an azido-containing functionality, which can be covalently conjugated in a second step with a complementary reaction partner, DBCO (dibenzocyclooctyne). We show that multimeric DBCO behaves quite differently than monoDBCO by imposing steric and topological constraints on RNAs. This generalizable strategy lays a solid foundation for developing clickable CRISPR off switches. We also show that this strategy works well for terminating CRISPR-Cas9-mediated genome editing in human cells.