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Light-mediated Reversible Modulation of the Mitogen-activated Protein Kinase Pathway during Cell Differentiation and Xenopus Embryonic Development

Light-mediated Reversible Modulation of the Mitogen-activated Protein Kinase Pathway during Cell Differentiation and Xenopus Embryonic Development

Published on: June 15, 2017

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

Takahiro Nakajima^1,2, 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

This summary is machine-generated.

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.

Keywords:

CRISPR–Cas9 Endogenous gene activation Optogenetics Photoswitch Red light

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Published on: August 4, 2021

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.