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Updated: Mar 22, 2026

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

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Published on: June 15, 2017

Optimized second-generation CRY2-CIB dimerizers and photoactivatable Cre recombinase.

Amir Taslimi¹, Brian Zoltowski², Jose G Miranda¹

¹Department of Pharmacology, University of Colorado School of Medicine, Aurora, Colorado, USA.

Nature Chemical Biology

|April 12, 2016

Summary

Researchers optimized the CRY2-CIB optogenetic system for better control of protein interactions. They developed a new photoactivatable Cre recombinase (PA-Cre2.0) with enhanced dynamic range for precise light-induced genetic manipulation.

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

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Spatiotemporal Control of Protein Activity through Optogenetic Allosteric Regulation

Spatiotemporal Control of Protein Activity through Optogenetic Allosteric Regulation

Published on: October 4, 2024

Area of Science:

Molecular Biology
Optogenetics
Plant Science

Background:

Arabidopsis thaliana cryptochrome 2 (AtCRY2) and CIB1 form a light-dependent dimerization system used in optogenetics.
Existing CRY2-CIB systems have limitations including size and dark interactions.

Purpose of the Study:

To optimize the CRY2-CIB system for improved function.
To identify smaller CRY2-CIB domains with reduced dark interaction and tunable signaling states.
To engineer enhanced photoactivatable tools for optogenetics.

Main Methods:

Characterization of minimal functional domains of CRY2 and CIB1.
Identification and analysis of mutations affecting AtCRY2 photocycle kinetics.
Engineering of a second-generation photoactivatable Cre recombinase (PA-Cre2.0) using a long-lived photocycle mutant.

Main Results:

Identified minimal functional CRY2 and CIB1 domains enabling light-dependent interaction.
Discovered signaling mutations in AtCRY2, implicating an α13-α14 turn motif in photocycle kinetics.
Developed PA-Cre2.0 with a five-fold improved dynamic range for robust recombination after brief light exposure.

Conclusions:

Optimized CRY2-CIB system offers enhanced control over protein interactions.
New mutations provide insights into CRY photocycle regulation and signaling.
PA-Cre2.0 represents a significant advancement in optogenetic tools for precise genetic engineering.