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Light-Controlled Fermentations for Microbial Chemical and Protein Production
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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
PubMed
Summary
This summary is machine-generated.

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.

Keywords:
Bacillus subtilisCcaSROptogeneticsSynthetic biologyTwo-component system

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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.