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

ACS Synthetic Biology
Shaobin Guo, Richard M Murray

Abstract

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.

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Citations

Nov 30, 2019·Nature Reviews. Genetics·Adam D SilvermanMichael C Jewett
Dec 17, 2019·Biotechnology and Bioengineering·Mohammad H Ayoubi-JoshaghaniRana Jahanban-Esfahlan
Aug 15, 2020·Frontiers in Bioengineering and Biotechnology·Nadanai Laohakunakorn
Jun 2, 2021·ACS Synthetic Biology·Pascal A PietersTom F A de Greef
Aug 7, 2021·Frontiers in Bioengineering and Biotechnology·Megan A McSweeney, Mark P Styczynski

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