Light-controllable RNA-protein devices for translational regulation of synthetic mRNAs in mammalian cells

Hideyuki Nakanishi; Tatsuyuki Yoshii; Shunsuke Kawasaki; Karin Hayashi; Keita Tsutsui; Choji Oki; Shinya Tsukiji; Hirohide Saito

doi:10.1016/j.chembiol.2021.01.002

Light-controllable RNA-protein devices for translational regulation of synthetic mRNAs in mammalian cells

Cell Chem Biol. 2021 May 20;28(5):662-674.e5. doi: 10.1016/j.chembiol.2021.01.002. Epub 2021 Jan 27.

Authors

Hideyuki Nakanishi¹, Tatsuyuki Yoshii², Shunsuke Kawasaki³, Karin Hayashi³, Keita Tsutsui⁴, Choji Oki⁵, Shinya Tsukiji⁶, Hirohide Saito⁷

Affiliations

¹ Department of Life Science Frontiers, Center for iPS Cell Research and Application, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan; Department of Biofunction Research, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University (TMDU), 2-3-10 Kanda-Surugadai, Chiyoda-ku, Tokyo 101-0062, Japan.
² Department of Life Science and Applied Chemistry, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan; PRESTO, Japan Science and Technology Agency (JST), 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan.
³ Department of Life Science Frontiers, Center for iPS Cell Research and Application, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
⁴ Department of Life Science and Applied Chemistry, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan.
⁵ Department of Nanopharmaceutical Sciences, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan.
⁶ Department of Life Science and Applied Chemistry, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan; Department of Nanopharmaceutical Sciences, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan.
⁷ Department of Life Science Frontiers, Center for iPS Cell Research and Application, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan. Electronic address: hirohide.saito@cira.kyoto-u.ac.jp.

PMID: 33508227
DOI: 10.1016/j.chembiol.2021.01.002

Abstract

The photo-regulation of transgene expression is one effective approach in mammalian synthetic biology due to its high spatial and temporal resolution. While DNAs are mainly used as vectors, modified RNAs (modRNAs) are also useful for medical applications of synthetic biology, because they can avoid insertional mutagenesis and immunogenicity. However, the optogenetic control of modRNA-delivered transgenes is much more difficult than that of DNA-delivered transgenes. Here, we develop two types of photo-controllable translational activation systems that are compatible with modRNAs. One is composed of a heterodimerization domain-fused split translational activator protein and a photocaged heterodimerizer. The other is composed of a destabilizing domain-fused translational activator protein and a photocaged stabilizer. The destabilized type can be used for not only translational activation but also translational repression of the modRNAs. These photo-controllable translation systems will expand the application of mammalian synthetic biology research.

Keywords: RNA binding protein; modified nucleoside; optogenetics; photocage; synthetic biology; synthetic mRNA; translation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

HeLa Cells
Humans
Light*
RNA, Messenger / biosynthesis*
RNA, Messenger / chemistry
RNA-Binding Proteins / chemistry
RNA-Binding Proteins / metabolism*
Tumor Cells, Cultured

Substances

RNA, Messenger
RNA-Binding Proteins