Sigma Factor-Mediated Tuning of Bacterial Cell-Free Synthetic Genetic Oscillators
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| Název: | Sigma Factor-Mediated Tuning of Bacterial Cell-Free Synthetic Genetic Oscillators |
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| Autoři: | Maaruthy Yelleswarapu, Ardjan J. van der Linden, Bob van Sluijs, Pascal A. Pieters, Emilien Dubuc, Tom F. A. de Greef, Wilhelm T. S. Huck |
| Zdroj: | ACS Synth Biol ACS Synthetic Biology |
| Informace o vydavateli: | American Chemical Society (ACS), 2018. |
| Rok vydání: | 2018 |
| Témata: | 0301 basic medicine, 0303 health sciences, Cell-Free System, competition-induced regulation, Bacterial, Sigma Factor, DNA-Directed RNA Polymerases, Gene Expression Regulation, Bacterial, DNA-Directed RNA Polymerases/genetics, Escherichia coli/genetics, 03 medical and health sciences, Gene Expression Regulation, Synthetic Biology/methods, cell-free systems, oscillator, Escherichia coli, Synthetic Biology, synthetic biology, Sigma Factor/genetics, Algorithms, sigma factors |
| 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. |
| Druh dokumentu: | Article Other literature type |
| Jazyk: | English |
| ISSN: | 2161-5063 |
| DOI: | 10.1021/acssynbio.8b00300 |
| Přístupová URL adresa: | https://pubs.acs.org/doi/pdf/10.1021/acssynbio.8b00300 https://pubmed.ncbi.nlm.nih.gov/30408412 http://www.ncbi.nlm.nih.gov/pubmed/30408412 https://europepmc.org/abstract/MED/30408412 https://pubs.acs.org/doi/10.1021/acssynbio.8b00300 https://research.tue.nl/en/publications/sigma-factor-mediated-tuning-of-bacterial-cell-free-synthetic-gen https://pure.tue.nl/ws/files/116746396/acssynbio.8b00300.pdf https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6305555 |
| Rights: | CC BY NC ND URL: http://pubs.acs.org/page/policy/authorchoice_ccbyncnd_termsofuse.html |
| Přístupové číslo: | edsair.doi.dedup.....ffe0f7fc2c50ca44b98aa6c41c28d85e |
| Databáze: | OpenAIRE |
Nájsť tento článok vo Web of Science | Abstrakt: | 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. |
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| ISSN: | 21615063 |
| DOI: | 10.1021/acssynbio.8b00300 |