Synthetic Mechanotransduction Using Engineered SynNotch Receptors

Cells can sense and interpret mechanical stimuli from their environments and neighbors, but the ability to engineer customized mechanosensing capabilities has remained a synthetic and mechanobiology challenge. Here, we introduce tension-tuned synthetic Notch (SynNotch) receptors that can be used to convert extracellular and intercellular forces into specifiable gene expression changes. By elevating the tension requirements of SynNotch activation, in combination with structure-guided mutagenesis, we designed a set of receptors with mechanical sensitivities spanning the physiologically relevant picoNewton (pN) range. Cells expressing these receptors can distinguish between varying tensile forces and respond by enacting customizable transcriptional programs. The synthetic utility of these tools is demonstrated by designing a decision-making circuit, through which fibroblasts can be made to differentiate into myoblasts upon stimulation with distinct tension magnitudes. Mechanobiological utility is also demonstrated by characterizing cell-generated forces transmitted between cells during Notch signaling. Overall, this work provides insight regarding how mechanically induced changes in protein structure can be used to transduce physical forces into biochemical signals. The system should facilitate the further programming and dissection of force-related phenomena in biological systems.