Cell-Free Characterization of Coherent Feed-Forward Loop-Based Synthetic Genetic Circuits

Pascal A. Pieters, Bryan L. Nathalia, Ardjan J. van der Linden, Peng Yin, Jongmin Kim (Corresponding author), Wilhelm T.S. Huck (Corresponding author), Tom F.A. de Greef (Corresponding author)

Onderzoeksoutput: Bijdrage aan tijdschriftTijdschriftartikelAcademicpeer review

2 Citaten (Scopus)
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Samenvatting

Regulatory pathways inside living cells employ feed-forward architectures to fulfill essential signal processing functions that aid in the interpretation of various types of inputs through noise-filtering, fold-change detection and adaptation. Although it has been demonstrated computationally that a coherent feed-forward loop (CFFL) can function as noise filter, a property essential to decoding complex temporal signals, this motif has not been extensively characterized experimentally or integrated into larger networks. Here we use post-transcriptional regulation to implement and characterize a synthetic CFFL in an Escherichia coli cell-free transcription-translation system and build larger composite feed-forward architectures. We employ microfluidic flow reactors to probe the response of the CFFL circuit using both persistent and short, noise-like inputs and analyze the influence of different circuit components on the steady-state and dynamics of the output. We demonstrate that our synthetic CFFL implementation can reliably repress background activity compared to a reference circuit, but displays low potential as a temporal filter, and validate these findings using a computational model. Our results offer practical insight into the putative noise-filtering behavior of CFFLs and show that this motif can be used to mitigate leakage and increase the fold-change of the output of synthetic genetic circuits.

Originele taal-2Engels
Pagina's (van-tot)1406-1416
Aantal pagina's11
TijdschriftACS Synthetic Biology
Volume10
Nummer van het tijdschrift6
DOI's
StatusGepubliceerd - 1 jun 2021

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Publisher Copyright:
© 2021 The Authors. Published by American Chemical Society.

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