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Engineering Transcriptional Regulator Effector Specificity Through Rational Design and Rapid Prototyping
Abstract The pursuit of circuits and metabolic path …
The pursuit of circuits and metabolic pathways of increasing complexity and robustness in synthetic biology will require engineering new regulatory tools. Feedback control based on relevant molecules, including toxic intermediates and environmental signals, would enable genetic circuits to react appropriately to changing conditions. In this work, computational protein design was used to create functional variants of qacR, a tetR family repressor, responsive to a new targeted effector. The modified repressors target vanillin, a growth-inhibiting small molecule found in lignocellulosic hydrolysates and other industrial processes. A computatio ally designed library was screened using an in vitro transcription-translation (TX-TL) system. Leads from the in vitro screen were characterized in vivo. Preliminary results demonstrate dose-dependent regulation of a downstream fluorescent reporter by vanillin. These repressor designs provide a starting point for the evolution of improved variants. We believe this process can serve as a framework for designing new sensors for other target compounds.
ng new sensors for other target compounds.  +
Authors Emmanuel L.C. de los Santos, Joseph T. Meyerowitz, Stephen L. Mayo, and Richard M. Murray  +
Funding Biomolecular Circuits for Rapid Detection and Response to Environmental Events + , Programmable Molecular Technology Initiative +
ID 2014g  +
Source Presented at Synthetic Biology: Engineering, Evolution and Design (SEED), 14-18 July 2014  +
Tag smmm14-seed  +
Title Engineering Transcriptional Regulator Effector Specificity Through Rational Design and Rapid Prototyping +
Type Conference Preprint  +
Categories Papers
Modification date
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15 May 2016 06:14:45  +
URL
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http://www.cds.caltech.edu/~murray/preprints/smmm14-seed.pdf  +
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Engineering Transcriptional Regulator Effector Specificity Through Rational Design and Rapid Prototyping + Title
 

 

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