Sigma Factor-Mediated Tuning of Bacterial Cell-Free Synthetic Genetic Oscillators
| Autor: | Emilien Dubuc, Wilhelm T. S. Huck, Ardjan J. van der Linden, Tom F. A. de Greef, Maaruthy Yelleswarapu, Pascal A. Pieters, Bob van Sluijs |
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| Přispěvatelé: | Synthetic Biology, Chemical Biology, Computational Biology |
| Jazyk: | angličtina |
| Předmět: |
0301 basic medicine
competition-induced regulation Computer science Biomedical Engineering Evolutionary algorithm Sigma Factor Model parameters Biochemistry Genetics and Molecular Biology (miscellaneous) 03 medical and health sciences Synthetic biology Sigma factor cell-free systems oscillator Escherichia coli Transcriptional regulation sigma factors Cell-Free System Kinetic model Bacterial DNA-Directed RNA Polymerases Gene Expression Regulation Bacterial General Medicine DNA-Directed RNA Polymerases/genetics Escherichia coli/genetics 030104 developmental biology Gene Expression Regulation Synthetic Biology/methods Synthetic Biology Sigma Factor/genetics Biological system Algorithms Research Article |
| Zdroj: | ACS Synthetic Biology ACS Synthetic Biology, 7(12), 2879-2887. American Chemical Society |
| ISSN: | 2161-5063 |
| DOI: | 10.1021/acssynbio.8b00300 |
| Popis: | Cell-free transcription-translation provides a simplified prototyping environment to rapidly design and study synthetic networks. Despite the presence of a well characterized toolbox of genetic elements, examples of genetic networks that exhibit complex temporal behavior are scarce. Here, we present a genetic oscillator implemented in an E. coli-based cell-free system under steady-state conditions using microfluidic flow reactors. The oscillator has an activator-repressor motif that utilizes the native transcriptional machinery of E. coli: the RNAP and its associated sigma factors. We optimized a kinetic model with experimental data using an evolutionary algorithm to quantify the key regulatory model parameters. The functional modulation of the RNAP was investigated by coupling two oscillators driven by competing sigma factors, allowing the modification of network properties by means of passive transcriptional regulation. |
| Databáze: | OpenAIRE |
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