A Red Light-Activatable Endogenous Gene Transcription System with Red-CPTS

Takahiro Nakajima; Yuto Kuwasaki; Shota Yamamoto; Takahiro Otabe; Moritoshi Sato

doi:10.1007/978-1-0716-4047-0_4

A Red Light-Activatable Endogenous Gene Transcription System with Red-CPTS

Methods Mol Biol. 2025:2840:45-55. doi: 10.1007/978-1-0716-4047-0_4.

Authors

Takahiro Nakajima^{1

2}, Yuto Kuwasaki², Shota Yamamoto², Takahiro Otabe^{1

2}, Moritoshi Sato^{3

4}

Affiliations

¹ Kanagawa Institute of Industrial Science and Technology, Kawasaki-shi, Kanagawa, Japan.
² Graduate School of Arts and Sciences, The University of Tokyo, Tokyo, Japan.
³ Kanagawa Institute of Industrial Science and Technology, Kawasaki-shi, Kanagawa, Japan. [email protected].
⁴ Graduate School of Arts and Sciences, The University of Tokyo, Tokyo, Japan. [email protected].

PMID: 39724343
DOI: 10.1007/978-1-0716-4047-0_4

Abstract

Red light penetrates deep into mammalian tissues and has low phototoxicity. We developed a red light-activatable photoswitch (MagRed) for deep tissue optogenetics. Using MagRed, we developed a red light-activatable endogenous gene transcription system (Red-CPTS) based on CRISPR-Cas9. Here we provide a detailed protocol for endogenous gene activation using Red-CPTS in cultured mammalian cells and living mice in vivo.

Keywords: CRISPR–Cas9; Endogenous gene activation; Optogenetics; Photoswitch; Red light.

MeSH terms

Animals
CRISPR-Cas Systems*
Humans
Light*
Mice
Optogenetics* / methods
Red Light
Transcription, Genetic* / radiation effects