Optogenetic tools can provide direct and programmable control of gene expression. Light-inducible recombinases, in particular, offer a powerful method for achieving precise spatiotemporal control of DNA modification. However, to-date this technology has been largely limited to eukaryotic systems. Here, we develop optogenetic recombinases for Escherichia coli that activate in response to blue light. Our approach uses a split recombinase coupled with photodimers, where blue light brings the split protein together to form a functional recombinase. We tested both Cre and Flp recombinases, Vivid and Magnet photodimers, and alternative protein split sites in our analysis. The optimal configuration, Opto-Cre-Vvd, exhibits strong blue light-responsive excision and low ambient light sensitivity. For this system we characterize the effect of light intensity and the temporal dynamics of light-induced recombination. These tools expand the microbial optogenetic toolbox, offering the potential for precise control of DNA excision with light-inducible recombinases in bacteria.

中文翻译：

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用于细菌光遗传学的光诱导重组酶。

光遗传学工具可以提供对基因表达的直接和可编程控制。特别是光诱导重组酶提供了一种强大的方法来实现对 DNA 修饰的精确时空控制。然而，迄今为止，这项技术在很大程度上仅限于真核系统。在这里，我们开发了大肠杆菌的光遗传学重组酶，可响应蓝光而激活。我们的方法使用与光二聚体结合的分裂重组酶，其中蓝光将分裂的蛋白质聚集在一起形成功能性重组酶。我们在分析中测试了 Cre 和 Flp 重组酶、Vivid 和 Magnet 光二聚体以及替代蛋白质分裂位点。最佳配置 Opto-Cre-Vvd 表现出强烈的蓝光响应切除和低环境光敏感性。对于这个系统，我们描述了光强度的影响和光诱导重组的时间动态。这些工具扩展了微生物光遗传学工具箱，为使用细菌中的光诱导重组酶精确控制 DNA 切除提供了潜力。