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Escherichia coli “Marionette” strains with 12 highly optimized small-molecule sensors

Escherichia coli “Marionette” strains with 12 highly optimized small-molecule sensors 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 cross-talk. 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Nature Chemical Biology Springer Journals

Escherichia coli “Marionette” strains with 12 highly optimized small-molecule sensors

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References (88)

Publisher
Springer Journals
Copyright
Copyright © 2018 by The Author(s), under exclusive licence to Springer Nature America, Inc.
Subject
Chemistry; Chemistry/Food Science, general; Biochemical Engineering; Biochemistry, general; Cell Biology; Bioorganic Chemistry
ISSN
1552-4450
eISSN
1552-4469
DOI
10.1038/s41589-018-0168-3
Publisher site
See Article on Publisher Site

Abstract

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 cross-talk. 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.

Journal

Nature Chemical BiologySpringer Journals

Published: Nov 26, 2018

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