Cell-Free Characterization of Coherent Feed-Forward Loop-Based Synthetic Genetic Circuits
SCIE
SCOPUS
- Title
- Cell-Free Characterization of Coherent Feed-Forward Loop-Based Synthetic Genetic Circuits
- Authors
- Pieters, Pascal A; Nathalia, Bryan L; van der Linden, Ardjan J; Yin, Peng; KIM, JONGMIN; Huck, Wilhelm TS; de Greef, Tom FA
- Date Issued
- 2021-06-18
- Publisher
- American Chemical Society
- Abstract
- 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.
- Keywords
- SMALL RNA; NETWORK; LOGIC; EXPRESSION; TOOLBOX; DYNAMICS
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/106813
- DOI
- 10.1021/acssynbio.1c00024
- ISSN
- 2161-5063
- Article Type
- Article
- Citation
- ACS SYNTHETIC BIOLOGY, vol. 10, no. 6, page. 1406 - 1416, 2021-06-18
- Files in This Item:
- There are no files associated with this item.
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.