| Summary: | Summary: 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. : Yamada et al. develop photoactivatable (PA)-Tet-OFF/ON systems for precise temporal and spatial control of cellular gene expression. The PA-Tet-OFF/ON can be applied to various cell types in vitro and in vivo, and their transcriptional activities are tightly regulated by blue light illumination and the small molecule Dox. Keywords: optogenetics, Tet system, Tet-OFF, Tet-ON, Cry2-CIB1
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