Regulatable and Modulable Background Expression Control in Prokaryotic Synthetic Circuits by Auxiliary Repressor Binding Sites

ACS Synthetic Biology
Davide Merulla, Jan Roelof van der Meer

Abstract

Expression control in synthetic genetic circuitry, for example, for construction of sensitive biosensors, is hampered by the lack of DNA parts that maintain ultralow background yet achieve high output upon signal integration by the cells. Here, we demonstrate how placement of auxiliary transcription factor binding sites within a regulatable promoter context can yield an important gain in signal-to-noise output ratios from prokaryotic biosensor circuits. As a proof of principle, we use the arsenite-responsive ArsR repressor protein from Escherichia coli and its cognate operator. Additional ArsR operators placed downstream of its target promoter can act as a transcription roadblock in a distance-dependent manner and reduce background expression of downstream-placed reporter genes. We show that the transcription roadblock functions both in cognate and heterologous promoter contexts. Secondary ArsR operators placed upstream of their promoter can also improve signal-to-noise output while maintaining effector dependency. Importantly, background control can be released through the addition of micromolar concentrations of arsenite. The ArsR-operator system thus provides a flexible system for additional gene expression control, which, g...Continue Reading

References

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Citations

Oct 19, 2016·Current Opinion in Microbiology·Robert W BradleyBaojun Wang
Feb 22, 2017·ACS Synthetic Biology·Yves BersetJan R van der Meer
May 17, 2018·Applied Microbiology and Biotechnology·Yun FangXianqiang Li
Jan 31, 2018·Journal of Industrial Microbiology & Biotechnology·Shuobo ShiHuimin Zhao
Dec 19, 2017·Biotechnology and Bioengineering·Xue-Feng ChenZhi-Gang Qian
Oct 1, 2020·Expert Opinion on Pharmacotherapy·Athina TrachalakiAthol U Wells
Mar 7, 2018·Proceedings of the National Academy of Sciences of the United States of America·Stephanie J DoongKristala L J Prather
Dec 1, 2017·Analytical and Bioanalytical Chemistry·Hyun Ju KimSang Jun Lee
Mar 27, 2019·Nature Chemical Biology·Xinyi WanBaojun Wang
Apr 13, 2021·World Journal of Microbiology & Biotechnology·Syed Azmal AliGurjeet Kaur
Sep 12, 2020·Ecotoxicology and Environmental Safety·Xuanyu WangShaopeng Chen
Aug 2, 2021·Metabolic Engineering·Ji Yeon LeeGyoo Yeol Jung

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