Sculpting at microscale: Constructing membraneless organelles in synthetic cells

Synthetic cells are engineered systems that mimic biological cells using basic components, offering insights into cell biology and enabling biotechnological advances. In living cell systems, membraneless organelles (MOs) play crucial roles and interact with membranes, making them essential for designing functional synthetic cells. As a result, understanding and applying model MOs is key to engineering complex synthetic cell systems, achieve basic cellular functions, and move towards specific applications. As one of the key governing mechanisms behind intercellular compartmentalization, liquid-liquid phase separation (LLPS), both associative and segregation, has garnered increased recognition for creating distinct internal environments in synthetic cells. However, effective compartmentalization, organization and membrane remodeling of synthetic cells through LLPS remains a significant challenge.

In Chapter 2, we present a microfluidic system for producing cell-mimicking containers. In Chapter 3, we investigate reversibly triggered condensation within our synthetic cell models. In Chapter 4, we turned our attention to coacervate interaction with the lipid bilayer membrane of liposome-based synthetic cells. In Chapter 5, we construct shells and scaffolds from solidified condensates and elucidate their potential applications for synthetic cells. Finally, in Chapter 6, we discuss the significance of this thesis, current advances and limitations of segregative and associative phase separation in synthetic cell study, as well as future opportunities for their use in applications.