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Construction of Incoherent Feedforward Loop Circuits in a Cell-Free System and in Cells

Guo, Shaobin and Murray, Richard M. (2019) Construction of Incoherent Feedforward Loop Circuits in a Cell-Free System and in Cells. ACS Synthetic Biology, 8 (3). pp. 606-610. ISSN 2161-5063. doi:10.1021/acssynbio.8b00493.

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Cells utilize transcriptional regulation networks to respond to environmental signals. Network motifs, such as feedforward loops, play essential roles in these regulatory networks. In this work, we construct two different functional and modular incoherent type 1 feedforward loop circuits in a cell-free transcription–translation system and in cells. With the help of mathematical modeling and the cell-free system, we can streamline the design–build–test cycles of the circuits, in which we characterize and optimize these circuits in vitro to confirm that they function as expected before implementing them in vivo. We show that the performance of these circuits from in vitro studies closely recapitulates those from in vivo experiments. We demonstrate that these feedforward loops show dynamic response and pulse-like behavior both in vitro and in vivo. These novel feedforward loop network motifs can be incorporated in more complicated biological circuits as detectors or responders.

Item Type:Article
Related URLs:
URLURL TypeDescription Information
Guo, Shaobin0000-0001-9736-4078
Murray, Richard M.0000-0002-5785-7481
Additional Information:© 2019 American Chemical Society. Received: November 25, 2018; Published: February 21, 2019. We would like to thank Clarmyra Hayes, Zachary Sun, Yutaka Hori, and Vipul Singhal for helpful discussion and suggestions. We would like to thank Murray lab members for useful suggestions. This work was supported by DARPA Living Foundries (Grant number: HR0011-12-C-0065). Author Contributions: S.G. and R.M.M. conceived the idea. S.G. designed the experiments, performed the experiments and subsequent data analysis, and wrote the manuscript. R.M.M. provided comments on the manuscript. The authors declare the following competing financial interest(s): R.M.M. has ownership in a company that commercializes the cell-free technology utilized in this paper. All the other authors claim no competing interest.
Funding AgencyGrant Number
Defense Advanced Research Projects Agency (DARPA)HR0011-12-C-0065
Subject Keywords:cell-free system, feedforward loop, biological circuit, prototyping, synthetic biology
Issue or Number:3
Record Number:CaltechAUTHORS:20190312-134813069
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Official Citation:Construction of Incoherent Feedforward Loop Circuits in a Cell-Free System and in Cells. Shaobin Guo and Richard M. Murray. ACS Synthetic Biology 2019 8 (3), 606-610. DOI: 10.1021/acssynbio.8b00493
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:93738
Deposited By: Tony Diaz
Deposited On:12 Mar 2019 21:14
Last Modified:16 Nov 2021 17:00

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