A tunable, rapid, and precise drug control of protein expression by combining transcriptional and post-translational regulation systems

作     者：Li Li Hongyi Meng Jianmin Zhang Yang Liu Qingjian Zou Yi Gao Huaqiang Yang Liangxue Lai Li Li;Hongyi Meng;Jianmin Zhang;Yang Liu;Qingjian Zou;Yi Gao;Huaqiang Yang;Liangxue Lai

作者机构：Department of Hepatobiliary Surgery IIGuangdong Provincial Research Center for Artificial Organ and Tissue EngineeringInstitute of Regenerative MedicineZhujiang HospitalSouthern Medical UniversityGuangzhou510282China CAS Key Laboratory of Regenerative BiologyGuangdong Provincial Key Laboratory of Stem Cell and Regenerative MedicineSouth China Institute for Stem Cell Biology and Regenerative MedicineGuangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou510530China School of Biotechnology and Health SciencesWuyi UniversityJiangmen529020China College of Animal ScienceSouth China Agricultural UniversityGuangzhou510642China 

出 版 物：《Journal of Genetics and Genomics》 (遗传学报（英文版）)

年 卷 期：2020年第47卷第11期

页      面：705-712页

核心收录：

学科分类：0710[理学-生物学] 1006[医学-中西医结合] 1001[医学-基础医学(可授医学、理学学位)] 10[医学] 100602[医学-中西医结合临床] 

基　　金：supported by the National Natural Science Foundation of China(81800555,81701580,and 31972926) the National Key R&D Program of China(2018YFC1106400) the Science and Technology Planning Project of Guangdong Province(2015B020229002) the Natural Science Foundation of Guangdong Province(2014A030312013 and 2018A030313128) 

主　　题：Protein abundance Tet-on 3G SMASh Gene expression CRISPR Small molecule 

摘      要：Rapid,precise,and tunable regulation of protein abundance would be significantly useful in a variety of biotechnologies and biomedical applications.Here,we describe a system that allows tunable and rapid drug control of gene expression for either gene activation or inactivation in mammalian cells.We construct the system by coupling Tet-on 3 G and small molecule-assisted shutoff systems,which can respectively induce transcriptional activation and protein degradation in the presence of corresponding small molecules.This dual-input drug inducer regulation system facilitates a bidirectional control of gene expression.The gene of interest can be precisely controlled by dual small molecules in a broad dynamic range of expression from overexpression to complete silence,allowing gene function study in a comprehensive expression profile.Our results reveal that the bidirectional control system enables sensitive dosage-and time-dependent regulation for either turn-on or shutoff of gene expression.We also apply this system for inducible genome editing and gene activation mediated by clustered regularly interspaced short palindromic repeats.The system provides an integrated platform for studying multiple biological processes by manipulating gene expression in a more flexible way.