Photo-Modulation of Gene-Editing Enzymes CRISPR/Cas9 with Bifunctional Small-Molecule Ligands  

作　　者：Yidan Zhang Yixin Zhang Lili Han Qiaoling Che Jiawei Tan Peng Zou Yiyun Chen 

机构地区：[1]State Key Laboratory of Chemical Biology,Shanghai Institute of Organic Chemistry,University of Chinese Academy of Sciences,Chinese Academy of Sciences,345 Lingling Road,Shanghai,200032 China [2]School of Chemistry and Material Sciences,Hangzhou Institute for Advanced Study,University of Chinese Academy of Sciences,1 Sub-lane Xiangshan,Hangzhou,Zhejiang,310024 China [3]School of Physical Science and Technology,ShanghaiTech University,100 Haike Road,Shanghai,201210 China

出　　处：《Chinese Journal of Chemistry》2023年第24期3639-3644,共6页中国化学（英文版）

基　　金：supported by the National Natural Science Foundation of China(22337005,22277133,91753126);the Youth Innovation Promotion Association(CAS 2023266);the CAS Interdisciplinary Innovation Team(JCTD-2020-16);the Program of Shanghai Academic/Technology Research Leader(21XD1424700).

摘　　要：The control of protein functions with light is valuable for spatiotemporal probing of biological systems.Current small-molecule photo-modulation methods include the light-induced uncaging of inhibitors and chromophore-assisted light inactivation with reactive oxygen species(ROS).However,the constant target protein expression results in inadequate photo-modulation efficiency,particularly for less potent inhibitors and chromophores.Herein,we report a novel bifunctional small-molecule ligands strategy to photo-modulate gene-editing enzymes CRISPR/Cas9.A coumarin-derived small-molecule ligand Bhc-BRD0539 is developed to uncage the active inhibitor upon light irradiation and to generate ROS in the Cas9 proximity for the dual inhibition of Cas9 activity.Our results highlight the synergistic photo-modulation with bifunctional small-molecule ligands,which offers a valuable addition to current CRISPR/Cas9 photo-modulation technologies and may extend to other protein classes.

关 键 词：Gene-editing Small-molecule ligands Enzyme modulation DNA cleavage PHOTOSWITCH Inhibitors 

分 类 号：Q814[生物学—生物工程]