Escherichia coli “Marionette” strains with 12 highly optimized small-molecule sensors
Name
285866v2.full.pdf
Description
Submitted version
Size
1.66 MB
Format
Adobe PDF
Checksum (MD5)
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Author(s)
Meyer, Adam J.
Segall-Shapiro, Thomas Hale
Glassey, Emerson
Zhang, Jing
Voigt, Christopher A.
Alternative Title
Escherichia coli “Marionette” strains with 12 highly optimized small-molecule sensors
Date Issued
November 2018
Journal
Nature Chemical Biology
Publisher
Springer Science and Business Media LLC
Citation
Meyer, Adam J. et al. "Escherichia coli “Marionette” strains with 12 highly optimized small-molecule sensors." Nature Chemical Biology 15 (November 2018): 196–204 © 2018 The Author(s)
Version
Original manuscript
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.
MIT Department
Massachusetts Institute of Technology. Synthetic Biology Center
Massachusetts Institute of Technology. Department of Biological Engineering
Terms of Use
Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1038/s41589-018-0168-3
