Optimization of Light-Inducible Gal4/UAS Gene Expression System in Mammalian Cells

Mayumi Yamada; Shinji C Nagasaki; Yusuke Suzuki; Yukinori Hirano; Itaru Imayoshi

doi:10.1016/j.isci.2020.101506

Optimization of Light-Inducible Gal4/UAS Gene Expression System in Mammalian Cells

iScience. 2020 Aug 27;23(9):101506. doi: 10.1016/j.isci.2020.101506. eCollection 2020 Sep 25.

Authors

Mayumi Yamada^{1

2

3

4}, Shinji C Nagasaki¹, Yusuke Suzuki^{1

2

4}, Yukinori Hirano^{4

5}, Itaru Imayoshi^{1

2

3

5

6}

Affiliations

¹ Research Center for Dynamic Living Systems, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan.
² Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto 606-8507, Japan.
³ World Premier International Research Initiative-Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto 606-8501, Japan.
⁴ Medical Innovation Center/SK Project, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
⁵ The Hakubi Center, Kyoto University, Kyoto 606-8302, Japan.
⁶ Japan Science and Technology Agency, Precursory Research for Embryonic Science and Technology, Saitama 332-0012, Japan.

Abstract

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.

Keywords: Cell Biology; Molecular Genetics; Optical Imaging.