Homologous Quorum Sensing Regulatory Circuit: A Dual-Input Genetic Controller for Modulating Quorum Sensing-Mediated Protein Expression in E. coli.,ACS Synthetic Biology

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Homologous Quorum Sensing Regulatory Circuit: A Dual-Input Genetic Controller for Modulating Quorum Sensing-Mediated Protein Expression in E. coli.
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2020-08-21 , DOI: 10.1021/acssynbio.0c00179
Pricila Hauk _{1,

2} , Kristina Stephens _{1,

2,

3} , Chelsea Virgile _{1,

2} , Eric VanArsdale _{1,

2,

3} , Alex Eli Pottash ₂ , John S Schardt ₂ , Steven M Jay ₂ , Herman O Sintim ₄ , William E Bentley _{1,

2,

3}

Affiliation

We developed a hybrid synthetic circuit that co-opts the genetic regulation of the native bacterial quorum sensing autoinducer-2 and imposes an extra external controller for maintaining tightly controlled gene expression. This dual-input genetic controller was mathematically modeled and, by design, can be operated in three modes: a constitutive mode that enables consistent and high levels of expression; a tightly repressed mode in which there is very little background expression; and an inducible mode in which concentrations of two signals (arabinose and autoinducer-2) determine the net amplification of the gene(s)-of-interest. We demonstrate the utility of the circuit for the controlled expression of human granulocyte macrophage colony stimulating factor in an engineered probiotic E. coli. This dual-input genetic controller is the first homologous AI-2 quorum sensing circuit that has the ability to be operated in three different modes. We believe it has the potential for wide-ranging biotechnological applications due its versatile features.

中文翻译：

同源群体感应调节电路：用于调节大肠杆菌中群体感应介导的蛋白质表达的双输入遗传控制器。

我们开发了一种混合合成电路，该电路结合了天然细菌群体感应自诱导剂 2 的遗传调控，并施加了一个额外的外部控制器来维持严格控制的基因表达。这种双输入基因控制器经过数学建模，按照设计，可以在三种模式下运行：一种能够实现一致和高水平表达的本构模式；一种严格压抑的模式，其中几乎没有背景表达；以及一种诱导模式，其中两种信号（阿拉伯糖和 autoinducer-2）的浓度决定目标基因的净扩增。我们证明了该电路在工程益生菌大肠杆菌中控制表达人粒细胞巨噬细胞集落刺激因子的效用. 这种双输入基因控制器是第一个能够在三种不同模式下运行的同源 AI-2 群体感应电路。我们相信它具有广泛的生物技术应用潜力，因为它具有多种功能。

更新日期：2020-10-17

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