A Red Light-Activatable Endogenous Gene Transcription System with Red-CPTS
Takahiro Nakajima1,2, Yuto Kuwasaki2, Shota Yamamoto2
1Kanagawa Institute of Industrial Science and Technology, Kawasaki-shi, Kanagawa, Japan.
Methods in Molecular Biology (Clifton, N.J.)
|December 26, 2024
Summary
We developed MagRed, a red light-activatable photoswitch, enabling deep tissue optogenetics. This system, Red-CPTS, uses CRISPR-Cas9 for precise gene transcription control in cells and living mice.
Area of Science:
- Optogenetics
- Molecular Biology
- Gene Editing
Background:
- Red light offers deep tissue penetration and low phototoxicity, ideal for biological applications.
- Existing optogenetic tools often lack efficiency in deep tissues.
- CRISPR-Cas9 technology provides precise genome manipulation capabilities.
Purpose of the Study:
- To develop a novel red light-activatable photoswitch (MagRed) for enhanced deep tissue optogenetics.
- To create a CRISPR-Cas9-based system (Red-CPTS) for red light-inducible endogenous gene transcription.
- To provide a detailed protocol for implementing Red-CPTS in mammalian cells and in vivo.
Main Methods:
- Development of the MagRed photoswitch.
- Integration of MagRed with CRISPR-Cas9 to form the Red-CPTS system.
- Protocol establishment for gene activation in cultured mammalian cells and live mice.
Main Results:
- Successful development of MagRed, a red light-activatable photoswitch.
- Creation of the Red-CPTS system for red light-controlled gene transcription.
- Demonstration of Red-CPTS efficacy in both cell cultures and in vivo mouse models.
Conclusions:
- MagRed and Red-CPTS offer a powerful new tool for deep tissue optogenetics.
- This system enables precise, red light-inducible control of endogenous gene transcription.
- The provided protocol facilitates the application of Red-CPTS in diverse research settings.
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