2. Academic Validation
  3. A tunable, rapid, and precise drug control of protein expression by combining transcriptional and post-translational regulation systems

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

  • J Genet Genomics. 2020 Nov 20;47(11):705-712. doi: 10.1016/j.jgg.2020.07.009.
Li Li 1,  Hongyi Meng 1,  Jianmin Zhang 1,  Yang Liu 2,  Qingjian Zou 3,  Yi Gao 4,  Huaqiang Yang 5,  Liangxue Lai 6
Affiliations

Affiliations

  • 1 Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, 510282, China.
  • 2 CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530, China.
  • 3 School of Biotechnology and Health Sciences, Wuyi University, Jiangmen, 529020, China. Electronic address: [email protected].
  • 4 Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, 510282, China. Electronic address: [email protected].
  • 5 College of Animal Science, South China Agricultural University, Guangzhou, 510642, China. Electronic address: [email protected].
  • 6 CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530, China; School of Biotechnology and Health Sciences, Wuyi University, Jiangmen, 529020, China. Electronic address: [email protected].
PMID: 33549479 DOI: 10.1016/j.jgg.2020.07.009
Abstract

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 3G 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.

Keywords

CRISPR; Gene expression; Protein abundance; SMASh; Small molecule; Tet-on 3G.

Figures
Products