Quorum-Sensing Crosstalk-Driven Synthetic Circuits: From Unimodality to Trimodality

• QS crosstalk is confirmed in LuxR/I and LasR/I by synthetic biology approaches
• Signal crosstalk causes shrinkage of bistable region while maintaining unimodality
• Promoter crosstalk reproducibly generates a complex trimodal response
• Model predictions and experimental validations of mutation-induced trimodality

SummaryWidespread quorum-sensing (QS) enables bacteria to communicate and plays a critical role in controlling bacterial virulence. However, effects of promiscuous QS crosstalk and its implications for gene regulation and cell decision-making remain largely unknown. Here we systematically studied the crosstalk between LuxR/I and LasR/I systems and found that QS crosstalk can be dissected into signal crosstalk and promoter crosstalk. Further investigations using synthetic positive feedback circuits revealed that signal crosstalk significantly decreases a circuit’s bistable potential while maintaining unimodality. Promoter crosstalk, however, reproducibly generates complex trimodal responses resulting from noise-induced state transitions and host-circuit interactions. A mathematical model that integrates the circuit’s nonlinearity, stochasticity, and host-circuit interactions was developed, and its predictions of conditions for trimodality were verified experimentally. Combining synthetic biology and mathematical modeling, this work sheds light on the complex behaviors emerging from QS crosstalk, which could be exploited for therapeutics and biotechnology.