Spatiotemporal Dynamic Assembly/Disassembly of Organelle‐Mimics Based on Intrinsically Disordered Protein‐Polymer Conjugates

Design of reversible organelle‐like microcompartments formed by liquid–liquid phase separation in cell‐mimicking entities has significantly advanced the bottom‐up construction of artificial eukaryotic cells. However, organizing the formation of artificial organelle architectures in a spatiotemporal manner within complex primitive compartments remains scarcely explored. In this work, thermoresponsive hybrid polypeptide‐polymer conjugates are rationally engineered and synthesized, resulting from the conjugation of an intrinsically disordered synthetic protein (IDP), namely elastin‐like polypeptide, and synthetic polymers (poly(ethylene glycol) and dextran) that are widely used as macromolecular crowding agents. Cell‐like constructs are built using droplet‐based microfluidics that are filled with such bioconjugates and an artificial cytoplasm system that is composed of specific polymers conjugated to the IDP. The distinct spatial organizations of two polypeptide‐polymer conjugates and the dynamic assembly and disassembly of polypeptide‐polymer coacervate droplets in response to temperature are studied in the cytomimetic protocells. Furthermore, a monoblock IDP with longer length is concurrently included with bioconjugates individually inside cytomimetic compartments. Both bioconjugates exhibit an identical surfactant‐like property, compartmentalizing the monoblock IDP coacervates via temperature control. These findings lay the foundation for developing hierarchically structured synthetic cells with interior organelle‐like structures which could be designed to localize in desired phase‐separated subcompartments.