Sensing and Reprogramming Surface Receptor Activation With Synthetic Transcriptional Circuits

ABSTRACT Cells rely on surface receptors to sense and initiate signalling cascades essential for numerous cellular processes, but engineering of synthetic genetic circuits to sense and rewire receptor activities for user‐defined cellular functions remains a challenge. Here we report a synthetic receptor‐signalling induced transcription (RESIT) circuit that enables sensing and reprogramming membrane‐localized receptor activation to pre‐defined transcriptional programs. The RESIT system is designed based on receptor activation mediated split protease complementation and release of membrane‐tethered synthetic transcriptional modules. We show that RESIT design is generally applicable to different transcriptional factors and various split viral proteases. This system is further engineered to probe Ca ²⁺ entry accompanied with PIEZO1 induction and T cell activation, to detect oncogenic receptor tyrosine kinase (RTK) activities and to assess Ras activation proximal to plasma membranes. The versatility of RESIT system is repurposed to actuate diverse therapeutic functions including apoptosis induction, target protein degradation and T cell activation in cells with high RTK activities. The modularity and versatility of RESIT highlight its promise for interrogating juxtamembrane biochemical signaling and rewiring receptor activation to therapeutic functions.