Bottom-Up Synthetic Biology for Artificial Cell Design: From Scaffold Materials to Functional Integration

Cells are extraordinary biochemical systems that have evolved over billions of years into highly sophisticated units of life. Bottom-up synthetic biology seeks to reconstruct cell-like systems from non-living molecular components, producing artificial cells that capture essential cellular features while bypassing the complexity and fragility of living organisms. This approach offers a unique perspective on the organizational principles underlying cellular life and provides a platform for diverse biotechnological applications. In this review, we survey recent advances in bottom-up artificial cell design, covering five principal scaffold materials including the newly prominent coacervate-based and hybrid hierarchical compartments, and four canonical functional categories: cascade metabolism, protein synthesis, division, and energy production. We further discuss the expanding application landscape spanning industrial biocatalysis, therapeutic protein delivery, biosensing, and origins of life research. Finally, we critically evaluate the key technical limitations currently facing the field, including module compatibility, operational stability, and regulatory challenges, and outline the directions that must be pursued to advance artificial cells toward practical realization.