Haller Daniel J, Castillo-Hair Sebastian M, Tabor Jeffrey J
Systems, Synthetic, and Physical Biology Program, Rice University, Houston, TX, USA.
Department of Bioengineering, Rice University, Houston, TX, USA.
Methods Mol Biol. 2025;2840:1-17. doi: 10.1007/978-1-0716-4047-0_1.
Optogenetics enables precise control of gene expression in a variety of organisms. We recently developed the first system for optogenetic control of transcription in Bacillus subtilis. This system is based on CcaSR, a light-responsive two-component regulatory system originally derived from Synechocystis PCC 6803. The so-called B. subtilis CcaSR v1.0 enables activation of gene expression with green light and deactivation with red. As a result, B. subtilis CcaSR v1.0 can be used to program gene expression with high quantitative, spatial, and temporal resolution. The expression levels of the CcaS light sensing histidine kinase and the CcaR response regulator are set by the addition of chemical inducers in B. subtilis CcaSR v1.0, enabling adjustment of the basal expression level and optimization of the magnitude of gene expression induction. In principle, B. subtilis CcaSR v1.0 should be compatible with expression of any target gene of interest. Here, we provide growth, strain engineering, and light treatment protocols for working with B. subtilis CcaSR.
光遗传学能够精确控制多种生物体中的基因表达。我们最近开发了首个用于枯草芽孢杆菌转录光遗传学控制的系统。该系统基于CcaSR，这是一种最初源自集胞藻PCC 6803的光响应双组分调节系统。所谓的枯草芽孢杆菌CcaSR v1.0能够通过绿光激活基因表达，并通过红光使其失活。因此，枯草芽孢杆菌CcaSR v1.0可用于以高定量、空间和时间分辨率对基因表达进行编程。在枯草芽孢杆菌CcaSR v1.0中，通过添加化学诱导剂来设定CcaS光感应组氨酸激酶和CcaR反应调节因子的表达水平，从而能够调整基础表达水平并优化基因表达诱导的幅度。原则上，枯草芽孢杆菌CcaSR v1.0应该与任何感兴趣的靶基因的表达兼容。在此，我们提供了使用枯草芽孢杆菌CcaSR的生长、菌株工程和光照处理方案。
