School of Biological and Health Systems Engineering, Arizona State University, Tempe, Arizona 85281, United States.
School of Mathematical and Statistical Sciences, Arizona State University, Tempe, Arizona 85281, United States.
ACS Synth Biol. 2021 May 21;10(5):1227-1236. doi: 10.1021/acssynbio.1c00041. Epub 2021 Apr 29.
Growth feedback, the inherent coupling between the synthetic gene circuit and the host cell growth, could significantly change the circuit behaviors. Previously, a diverse array of emergent behaviors, such as growth bistability, enhanced ultrasensitivity, and topology-dependent memory loss, were reported to be induced by growth feedback. However, the influence of the growth feedback on the circuit functions remains underexplored. Here, we reported an unexpected damped oscillatory behavior of a self-activation gene circuit induced by nutrient-modulating growth feedback. Specifically, after dilution of the activated self-activation switch into the fresh medium with moderate nutrients, its gene expression first decreases as the cell grows and then shows a significant overshoot before it reaches the steady state, leading to damped oscillation dynamics. Fitting the data with a coarse-grained model suggests a nonmonotonic growth-rate regulation on gene production rate. The underlying mechanism of the oscillation was demonstrated by a molecular mathematical model, which includes the ribosome allocation toward gene production, cell growth, and cell maintenance. Interestingly, the model predicted a counterintuitive dependence of oscillation amplitude on the nutrition level, where the highest peak was found in the medium with moderate nutrients, but was not observed in rich nutrients. We experimentally verified this prediction by tuning the nutrient level in the culture medium. We did not observe significant oscillatory behavior for the toggle switch, suggesting that the emergence of damped oscillatory behavior depends on circuit network topology. Our results demonstrated a new nonlinear emergent behavior mediated by growth feedback, which depends on the ribosome allocation between gene circuit and cell growth.
生长反馈,即合成基因电路与宿主细胞生长之间固有的耦合,会显著改变电路行为。此前,研究报道了多种新兴行为,如生长双稳态、增强的超敏性和拓扑依赖性记忆丢失,这些行为都是由生长反馈引起的。然而,生长反馈对电路功能的影响仍未得到充分探索。在这里,我们报道了一种由营养物质调节的生长反馈引起的自激活基因电路的出乎意料的衰减振荡行为。具体来说,在将激活的自激活开关稀释到含有适量营养物质的新鲜培养基中后,其基因表达首先随着细胞的生长而降低,然后在达到稳定状态之前出现明显的过冲,导致衰减振荡动力学。用粗粒化模型拟合数据表明,基因产生率的生长速率调节是非单调的。通过包括核糖体在基因产生、细胞生长和细胞维持方面的分配的分子数学模型,证明了振荡的潜在机制。有趣的是,该模型预测了振荡幅度与营养水平的反直觉依赖性,在中等营养物质的培养基中发现了最高峰值,但在丰富的营养物质中没有观察到。我们通过在培养基中调整营养水平实验验证了这一预测。我们没有观察到 toggle switch 的显著振荡行为,这表明衰减振荡行为的出现取决于电路网络拓扑。我们的研究结果表明,生长反馈介导的新的非线性新兴行为取决于核糖体在基因电路和细胞生长之间的分配。