Sigma Factor-Mediated Tuning of Bacterial Cell-Free Synthetic Genetic Oscillators.,ACS Synthetic Biology

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Sigma Factor-Mediated Tuning of Bacterial Cell-Free Synthetic Genetic Oscillators.
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-11-19 , DOI: 10.1021/acssynbio.8b00300
Maaruthy Yelleswarapu ₁ , Ardjan J van der Linden ₂ , Bob van Sluijs ₁ , Pascal A Pieters ₂ , Emilien Dubuc ₂ , Tom F A de Greef ₂ , Wilhelm T S Huck ₁

Affiliation

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.

中文翻译：

细菌无细胞合成遗传振荡器的 Sigma 因子介导的调谐。

无细胞转录-翻译为快速设计和研究合成网络提供了简化的原型设计环境。尽管存在一个特征明确的遗传元素工具箱，但表现出复杂时间行为的遗传网络的例子却很少。在这里，我们提出了一种在基于大肠杆菌的无细胞系统中使用微流体流动反应器在稳态条件下实现的遗传振荡器。振荡器有一个激活-抑制基序，它利用大肠杆菌的天然转录机制：RNAP 及其相关的 sigma 因子。我们使用进化算法优化了具有实验数据的动力学模型，以量化关键的调节模型参数。RNAP 的功能调制是通过耦合由竞争 sigma 因子驱动的两个振荡器来研究的，

更新日期：2018-11-08

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