Design of orthogonal genetic switches based on a crosstalk map of sigmas, anti-sigmas, and promoters.
Rhodius VA, Segall-Shapiro TH, Sharon BD, Ghodasara A, Orlova E, Tabakh H, Burkhardt DH, Clancy K, Peterson TC, Gross CA, Voigt CA
Mol Syst Biol. 2013 Oct 29;9:702. doi: 10.1038/msb.2013.58.
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PubMed
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Article
Using a combination of bioinformatics and functional assays, Christopher Voigt’s lab has identified a number of ECF σ factors, σ promoters, and anti-σ factors that are functional in E. coli. A subset of these elements act orthogonally to one another, and can be used to build orthogonal genetic switches.
This publication from the Voigt lab describes the identification, synthesis, and characterization of 86 ECF σ factors, their promoters, and 62 anti-σs. Fifty-two ECF σ factors were shown to be highly active in E. coli when paired with 24 promoters. Of these, a set of 20 orthogonal σ: promoter pairs was identified for use in building genetic programs. Similarly, 25 anti-σ factors were shown to strongly repress the activity of an active ECF σ factor, and a subset of 12 orthogonal pairs was identified.
The Voigt lab has deposited 52 pVRa plasmids encoding active ECF σ factors, 24 pVRb plasmids encoding σ promoters, 25 pVRc plasmids encoding anti-σ factors, and an accessory plasmid that produces T7 RNA polymerase (pN565).