Escherichia coli "Marionette" strains with 12 highly optimized small-molecule sensors
Escherichia coli "Marionette" strains with 12 highly optimized small-molecule sensors
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DOI：
10.1038/s41589-018-0168-3
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发表时间：
2019-02-01
影响因子：
14.8
通讯作者：
Voigt, Christopher A.
中科院分区：
文献类型：
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作者：
Meyer, Adam J.;Segall-Shapiro, Thomas H.;Voigt, Christopher A.
细胞过程由许多基因进行，它们的研究和优化需要多个杠杆，通过这些杠杆可以独立控制它们。最常见的方法是通过对小分子做出反应的遗传编码传感器。然而，这些传感器通常是次优的，表现出高背景表达和低动态范围。此外，在一个小区中使用多个传感器受到串扰和蜂窝资源负担的限制。在这里，我们开发了一种定向进化策略，可以同时选择更低的背景、高动态范围、更高的灵敏度和低串扰。这被应用于产生一组12个高性能传感器，其表现出> 100倍的诱导，具有低背景和交叉反应性。这些结合在一起，在大肠杆菌的基因组中建立了一个单一的“传感器阵列”。coli MG 1655（野生型）、DH10B（克隆）和BL 21（蛋白表达）。这些“木偶”菌株允许使用12种小分子诱导剂独立控制基因表达。
Cellular processes are carried out by many genes, and their study and optimization requires multiple levers by which they can be independently controlled. The most common method is via a genetically encoded sensor that responds to a small molecule. However, these sensors are often suboptimal, exhibiting high background expression and low dynamic range. Further, using multiple sensors in one cell is limited by cross-talk and the taxing of cellular resources. Here, we have developed a directed evolution strategy to simultaneously select for lower background, high dynamic range, increased sensitivity, and low crosstalk. This is applied to generate a set of 12 high-performance sensors that exhibit > 100-fold induction with low background and cross-reactivity. These are combined to build a single "sensor array" in the genomes of E. coli MG1655 (wild-type), DH10B (cloning), and BL21 (protein expression). These "Marionette" strains allow for the independent control of gene expression using 12 small-molecule inducers.