Optimization of Light-Inducible Gal4/UAS Gene Expression System in Mammalian Cells
Mayumi Yamada,
Shinji C. Nagasaki,
Yusuke Suzuki,
Yukinori Hirano,
Itaru Imayoshi
Affiliations
Mayumi Yamada
Research Center for Dynamic Living Systems, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan; Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto 606-8507, Japan; World Premier International Research Initiative–Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto 606-8501, Japan; Medical Innovation Center/SK Project, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan
Shinji C. Nagasaki
Research Center for Dynamic Living Systems, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan
Yusuke Suzuki
Research Center for Dynamic Living Systems, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan; Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto 606-8507, Japan; Medical Innovation Center/SK Project, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan
Yukinori Hirano
Medical Innovation Center/SK Project, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan; The Hakubi Center, Kyoto University, Kyoto 606-8302, Japan
Itaru Imayoshi
Research Center for Dynamic Living Systems, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan; Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto 606-8507, Japan; World Premier International Research Initiative–Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto 606-8501, Japan; The Hakubi Center, Kyoto University, Kyoto 606-8302, Japan; Japan Science and Technology Agency, Precursory Research for Embryonic Science and Technology, Saitama 332-0012, Japan; Corresponding author
Summary: Light-inducible gene expression systems represent powerful methods for studying the functional roles of dynamic gene expression. Here, we developed an optimized light-inducible Gal4/UAS gene expression system for mammalian cells. We designed photoactivatable (PA)-Gal4 transcriptional activators based on the concept of split transcription factors, in which light-dependent interactions between Cry2-CIB1 PA-protein interaction modules can reconstitute a split Gal4 DNA-binding domain and p65 transcription activation domain. We developed a set of PA-Gal4 transcriptional activators (PA-Gal4cc), which differ in terms of induced gene expression levels following pulsed or prolonged light exposure, and which have different activation/deactivation kinetics. These systems offer optogenetic tools for the precise manipulation of gene expression at fine spatiotemporal resolution in mammalian cells.
