期刊
NATURE CHEMICAL BIOLOGY
卷 15, 期 2, 页码 196-+出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/s41589-018-0168-3
关键词
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资金
- US Office of Naval Research Multidisciplinary University Research Initiative [N00014-16-1-2388]
- Koch Institute from the National Cancer Institute [P30-CA14051]
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.
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