RRC ID 54175
Author Yamada M, Suzuki Y, Nagasaki SC, Okuno H, Imayoshi I.
Title Light Control of the Tet Gene Expression System in Mammalian Cells.
Journal Cell Rep
Abstract Gene expression and its network structure are dynamically altered in multicellular systems during morphological, functional, and pathological changes. To precisely analyze the functional roles of dynamic gene expression changes, tools that manipulate gene expression at fine spatiotemporal resolution are needed. The tetracycline (Tet)-controlled gene expression system is a reliable drug-inducible method, and it is used widely in many mammalian cultured cells and model organisms. Here, we develop a photoactivatable (PA)-Tet-OFF/ON system for precise temporal control of gene expression at single-cell resolution. By integrating the cryptochrome 2-cryptochrome-interacting basic helix-loop-helix 1 (Cry2-CIB1) light-inducible binding switch, expression of the gene of interest is tightly regulated under the control of light illumination and drug application in our PA-Tet-OFF/ON system. This system has a large dynamic range of downstream gene expression and rapid activation/deactivation kinetics. We also demonstrate the optogenetic regulation of exogenous gene expression in vivo, such as in developing and adult mouse brains.
Volume 25(2)
Pages 487-500.e6
Published 2018-10-9
DOI 10.1016/j.celrep.2018.09.026
PII S2211-1247(18)31456-6
PMID 30304687
MeSH Animals Arabidopsis Proteins / genetics Arabidopsis Proteins / metabolism Basic Helix-Loop-Helix Transcription Factors / genetics Basic Helix-Loop-Helix Transcription Factors / metabolism Cells, Cultured Cryptochromes / genetics Cryptochromes / metabolism Female Gene Expression Regulation / drug effects* Light* Male Mice Mice, Inbred ICR Mice, Transgenic Neurons / drug effects Neurons / metabolism* Neurons / radiation effects Optogenetics / methods* Protein Synthesis Inhibitors / pharmacology* Tetracycline / pharmacology*
IF 8.109
Times Cited 13
Mice RBRC02340