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Red Light Responsive Cre Recombinase for Bacterial Optogenetics.

Fereshteh Jafarbeglou^1,2, Mary J Dunlop^1,2

¹Biomedical Engineering, Boston University, Boston, Massachusetts 02215, United States.

ACS Synthetic Biology

|November 19, 2024

Summary

We developed OptoCre-REDMAP, a red light-activated system for precise control in microbial engineering. This tool enables advanced synthetic biology applications with improved light penetration and orthogonality to blue light systems.

Keywords:

Cre recombinase bacterial optogenetics multichromatic control red light

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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:

Synthetic Biology
Microbial Engineering
Optogenetics

Background:

Optogenetic tools offer tunable control in microbial engineering.
Current systems primarily use blue light, limiting multichromatic applications.
Red light offers benefits like deeper penetration and reduced toxicity.

Purpose of the Study:

Introduce OptoCre-REDMAP, a red light-inducible Cre recombinase system for *Escherichia coli*.
Enable precise control over gene expression and DNA excision using red light.
Advance multichromatic control in microbial synthetic biology.

Main Methods:

Harnessed plant photoreceptors (PhyA, FHY1) and split Cre recombinase.
Optimized Cre start codon and characterized induction levels.
Tested sensitivity to light intensity, duration, and ambient conditions.

Main Results:

Achieved minimal dark expression and rapid activation (within 4h) with red light.
Demonstrated OptoCre-REDMAP's reliability and flexibility across various conditions.
Showcased orthogonal control with blue light systems (OptoCre-VVD) and superior red light penetration.

Conclusions:

OptoCre-REDMAP provides robust, selective red light control for microbial applications.
The system is suitable for advanced synthetic biology requiring precise multichromatic control.
Red light offers advantages over blue light for specific microbial engineering tasks.