Generalized Biosensing and Signaling Reprogramming Using Circular RNA-Protein Circuit-Mediated Cargo Release

Real-time monitoring and dynamic modulation of biological signals are crucial for understanding and reprogramming complex biological processes. However, modular sensing and actuation systems that respond to diverse signals remain largely unexplored. Here, we engineer a generalized biosensing and signaling reprogramming platform, circular RNA-protein circuit-mediated cargo release (CRMCR), enabling fast detection and precise rewiring of biological signals in a multicellular system. CRMCR is designed based on target signal-activated expression of a circular RNA scaffold, which mediates complementation of a split protease and facilitates the release of the reporter protein retained in the endoplasmic reticulum for signal production. We demonstrate that this platform allows detection of different biological signals including immunosuppressive factors, cytokines, and tumor-associated growth factors, with high sensitivity and fast response. Moreover, this design is further engineered as a reprogramming platform, allowing dynamic sensing and autonomous modulation of T cell activity in a multicellular system. This multifunctional platform provides a new direction for scalable detection and modulation of signals in a variety of biological scenarios.