Synthetic Tunable Amplifying Buffer Circuit in E. coli
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KayzadACS14(1).pdf
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Author(s)
Nilgiriwala, Kayzad
Jimenez Zarco, Jose I.
Rivera-Ortiz, Phillip Michael
Del Vecchio, Domitilla
Date Issued
October 2014
Journal
ACS Synthetic Biology
Publisher
American Chemical Society (ACS)
Citation
Nilgiriwala, Kayzad Soli, José Jiménez, Phillip Michael Rivera, and Domitilla Del Vecchio. “Synthetic Tunable Amplifying Buffer Circuit in E. Coli.” ACS Synthetic Biology 4, no. 5 (October 21, 2014): 577–584.
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Author's final manuscript
Abstract
While predictable design of a genetic circuit's output is a major goal of synthetic biology, it remains a significant challenge because DNA binding sites in the cell affect the concentration of available transcription factors (TF). To mitigate this problem, we propose to use a TF that results from the (reversible) phosphorylation of protein substrate as a circuit's output. We demonstrate that by comparatively increasing the amounts of substrate and phosphatase, the TF concentration becomes robust to the presence of DNA binding sites and can be kept at a desired value. The circuit's input/output gain can, in turn, be tuned by changing the relative amounts of the substrate and phosphatase, realizing an amplifying buffer circuit with tunable gain. In our experiments in E. coli, we employ phospho-NRI as the output TF, phosphorylated by the NRII kinase, and dephosphorylated by the NRII phosphatase. Amplifying buffer circuits such as ours could be used to insulate a circuit's output from the context, bringing synthetic biology one step closer to modular design.
MIT Department
Massachusetts Institute of Technology. Department of Mechanical Engineering
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DOI of Published Version
https://doi.org/10.1021/SB5002533
