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Optogenetic Control of B. subtilis Gene Expression Using the CcaSR System.
Daniel J Haller1, Sebastian M Castillo-Hair2,3, Jeffrey J Tabor4,5,6,7
1Systems, Synthetic, and Physical Biology Program, Rice University, Houston, TX, USA.
Methods in Molecular Biology (Clifton, N.J.)
|December 26, 2024
Summary
Researchers developed a novel optogenetic system for Bacillus subtilis, enabling precise control over gene expression using light. This system offers high-resolution programming of genetic circuits with green and red light stimuli.
Area of Science:
- Synthetic Biology
- Microbiology
- Molecular Biology
Background:
- Optogenetics offers precise control over biological processes.
- Controlling gene expression in Bacillus subtilis is crucial for various applications.
- Existing methods for gene expression control in B. subtilis have limitations.
Purpose of the Study:
- To develop and characterize the first optogenetic system for transcriptional control in Bacillus subtilis.
- To enable light-inducible and light-repressible gene expression with high spatiotemporal resolution.
- To provide protocols for utilizing this new system in B. subtilis.
Main Methods:
- Adaptation of the CcaSR two-component system from Synechocystis PCC 6803 for B. subtilis.
- Engineering of B. subtilis strains to express CcaS (histidine kinase) and CcaR (response regulator).
- Development of protocols for bacterial growth, strain engineering, and light-based induction/repression.
Main Results:
- Successful implementation of the B. subtilis CcaSR v1.0 system for light-controlled gene expression.
- Demonstration of gene expression activation with green light and deactivation with red light.
- Chemical inducers allow tuning of basal expression levels and induction magnitude.
Conclusions:
- The B. subtilis CcaSR v1.0 system provides a powerful tool for precise, light-mediated control of gene expression.
- This system is compatible with various target genes, expanding synthetic biology applications in B. subtilis.
- The provided protocols facilitate the adoption and use of optogenetic control in this important bacterial species.
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