Design of orthogonal genetic switches based on a crosstalk map of σs, anti-σs, and promoters
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Rhodius-2013-Design of orthogonal.pdf
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Author(s)
Rhodius, Virgil A.
Sharon, Brian D.
Orlova, Ekaterina
Tabakh, Hannah
Burkhardt, David H.
Clancy, Kevin
Peterson, Todd C.
Gross, Carol A.
Segall-Shapiro, Thomas H.
Ghodasara, Amar Navin
Date Issued
October 2013
Journal
Molecular Systems Biology
Publisher
Nature Publishing Group
Citation
Rhodius, Virgil A, Thomas H Segall-Shapiro, Brian D Sharon, Amar Ghodasara, Ekaterina Orlova, Hannah Tabakh, David H Burkhardt, et al. “Design of orthogonal genetic switches based on a crosstalk map of σs, anti-σs, and promoters.” Molecular Systems Biology 9 (October 29, 2013). Copyright © 2013 EMBO and Macmillan Publishers Limited
Version
Final published version
Abstract
Cells react to their environment through gene regulatory networks. Network integrity requires minimization of undesired crosstalk between their biomolecules. Similar constraints also limit the use of regulators when building synthetic circuits for engineering applications. Here, we mapped the promoter specificities of extracytoplasmic function (ECF) σs as well as the specificity of their interaction with anti‐σs. DNA synthesis was used to build 86 ECF σs (two from every subgroup), their promoters, and 62 anti‐σs identified from the genomes of diverse bacteria. A subset of 20 σs and promoters were found to be highly orthogonal to each other. This set can be increased by combining the −35 and −10 binding domains from different subgroups to build chimeras that target sequences unrepresented in any subgroup. The orthogonal σs, anti‐σs, and promoters were used to build synthetic genetic switches in Escherichia coli. This represents a genome‐scale resource of the properties of ECF σs and a resource for synthetic biology, where this set of well‐characterized regulatory parts will enable the construction of sophisticated gene expression programs.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
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DOI of Published Version
http://dx.doi.org/10.1038/msb.2013.58
