Spontaneous One-Pot Layer-by-Layer Assembly Enabling Programmable Protocell Communication

Designing artificial cell-like systems that reproduce the organizational and communicative complexity of living cells remains a major challenge in synthetic biology and materials science. Protocells, particularly coacervate-based microdroplets, offer powerful platforms for investigating compartmentalization, biochemical communication, and emergent collective behaviors. Yet, their lack of stable, multifunctional membranes severely limits their potential, as conventional interface-assembly methods typically allow only single-enzyme functionalization, leading to restricted morphological control and short-lived synthetic cell–cell communication. This project aims to overcome these limitations by developing a spontaneous one-pot layer-by-layer (SOP-LbL) assembly nanotechnology for the spatially programmable construction of multilayer enzyme architectures on protocell membranes. By integrating the applicant's expertise in polymer chemistry and materials science—especially in SOP-LbL assembly—with the host group's pioneering advances in protocell engineering, this interdisciplinary research will establish a robust and versatile strategy to create stable and functionally dynamic protocells. The resulting protocell populations will be capable of multi-round, programmable chemical communication, thereby emulating essential features of natural cellular communities. The project outcomes will generate fundamental insights into adaptive artificial cell design and provide a foundation for future applications in biomedical engineering and synthetic biology.