A modular toolset for electrogenetics
A modular toolset for electrogenetics
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DOI：
10.1101/2021.09.10.459750
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发表时间：
2021-09
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通讯作者：
J. Lawrence;Y. Yin;P. Bombelli;A. Scarampi;M. Storch;L. Wey;A. Climent-Catala;G. Baldwin;D. O’Hare;C. Howe;J. Zhang;T. Ouldridge;R. Ledesma‐Amaro
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文献类型：
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作者：
J. Lawrence;Y. Yin;P. Bombelli;A. Scarampi;M. Storch;L. Wey;A. Climent-Catala;G. Baldwin;D. O’Hare;C. Howe;J. Zhang;T. Ouldridge;R. Ledesma‐Amaro
合成生物学研究及其工业应用依赖于基因表达的确定性时空控制。最近，基因表达的电化学控制已被证明在电遗传系统（氧化还原响应的启动子一起使用的氧化还原诱导剂和电极），允许直接集成的电子与复杂的生物过程的各种新的应用。然而，电致系统的使用受到活性差、可调谐性和标准化的限制。在这里，我们已经开发了各种遗传和电化学工具，促进设计和大大提高电生系统的性能。我们开发了一个强大的，单向的，氧化还原反应的启动子，然后推导出一个突变的启动子库与频谱的优势。然后，我们用这些部分构建了遗传电路，并证明了它们被多种氧化还原分子激活。最后，我们证明了电化学激活的基因表达在有氧条件下，利用一种新的，模块化的生物电化学装置。该工具集为研究人员提供了为特定应用设计和构建优化的电遗传系统所需的所有元素。
Synthetic biology research and its industrial applications rely on the deterministic spatiotemporal control of gene expression. Recently, electrochemical control of gene expression has been demonstrated in electrogenetic systems (redox-responsive promoters used alongside redox inducers and an electrode), allowing for the direct integration of electronics with complex biological processes for a variety of new applications. However, the use of electrogenetic systems is limited by poor activity, tunability and standardisation. Here, we have developed a variety of genetic and electrochemical tools that facilitate the design and vastly improve the performance of electrogenetic systems. We developed a strong, unidirectional, redox-responsive promoter before deriving a mutant promoter library with a spectrum of strengths. We then constructed genetic circuits with these parts and demonstrated their activation by multiple classes of redox molecules. Finally, we demonstrated electrochemical activation of gene expression in aerobic conditions utilising a novel, modular bioelectrochemical device. This toolset provides researchers with all the elements needed to design and build optimised electrogenetic systems for specific applications.