Multiplexing Engineered Receptors for Multiparametric Evaluation of Environmental Ligands,ACS Synthetic Biology

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Multiplexing Engineered Receptors for Multiparametric Evaluation of Environmental Ligands
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2017-08-23 00:00:00 , DOI: 10.1021/acssynbio.6b00279
Rachel M. Hartfield ₁ , Kelly A. Schwarz ₁ , Joseph J. Muldoon _{1,

2} , Neda Bagheri _{1,

2,

3,

4,

5} , Joshua N. Leonard _{1,

2,

3,

4,

5}

Affiliation

Engineered cell-based therapies comprise a promising, emerging biomedical technology. Broad utilization of this strategy will require new approaches for implementing sophisticated functional programs, such as sensing and responding to the environment in a defined fashion. Toward this goal, we investigated whether our self-contained receptor and signal transduction system (MESA) could be multiplexed to evaluate extracellular cues, with a focus on elucidating principles governing the integration of such engineered components. We first developed a set of hybrid promoters that exhibited AND gate activation by two transcription factors. We then evaluated these promoters when paired with two MESA receptors and various ligand combinations. Unexpectedly, although the multiplexed system exhibited distinct responses to ligands applied individually and in combination, the same synergy was not observed as when promoters were characterized with soluble transcription factors. Therefore, we developed a mechanistic computational model leveraging these observations, to both improve our understanding of how the receptors and promoters interface and to guide the design and implementation of future systems. Notably, the model explicitly accounts for the impact of intercellular variation on system characterization and performance. Model analysis identified key factors that affect the current receptors and promoters, and enabled an in silico exploration of potential modifications that inform the design of improved logic gates and their robustness to intercellular variation. Ultimately, this quantitative design-driven approach may guide the use and multiplexing of synthetic receptors for diverse custom biological functions beyond the case study considered here.

中文翻译：

用于环境配体多参数评估的多路复用工程受体

基于细胞的工程疗法包括有前途的新兴生物医学技术。广泛使用此策略将需要新的方法来实施复杂的功能程序，例如以定义的方式感知和响应环境。为了实现这一目标，我们研究了是否可以将我们自成一体的受体和信号转导系统（MESA）进行多路复用以评估细胞外信号，重点是阐明控制此类工程化组件集成的原理。我们首先开发了一组杂交启动子，它们通过两个转录因子表现出AND门激活。然后，当与两个MESA受体和各种配体组合配对时，我们评估了这些启动子。不料，尽管多重系统对单独和组合使用的配体表现出不同的响应，但与使用可溶性转录因子表征启动子时未观察到相同的协同作用。因此，我们利用这些观察结果开发了一种机械的计算模型，以增进我们对受体和启动子如何相互作用的理解，并指导未来系统的设计和实施。值得注意的是，该模型明确说明了细胞间变异对系统表征和性能的影响。模型分析确定了影响当前受体和启动子的关键因素，并实现了我们利用这些观察结果开发了一种机械的计算模型，以增进我们对受体和启动子如何相互作用的理解，并指导未来系统的设计和实施。值得注意的是，该模型明确说明了细胞间变异对系统表征和性能的影响。模型分析确定了影响当前受体和启动子的关键因素，并实现了我们利用这些观察结果开发了一种机械的计算模型，以增进我们对受体和启动子如何相互作用的理解，并指导未来系统的设计和实施。值得注意的是，该模型明确说明了细胞间变异对系统表征和性能的影响。模型分析确定了影响当前受体和启动子的关键因素，并实现了在计算机上探索潜在的修饰，这些修饰可以告知改进的逻辑门设计及其对细胞间变异的鲁棒性。最终，这种定量设计驱动的方法可能会指导合成受体在多种自定义生物学功能中的使用和复用，而不是此处考虑的案例研究。

更新日期：2017-08-23

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