Abstract:
Membrane-less compartments formed via liquid−
liquid phase separation (LLPS) are regulated dynamically via
enzyme reactions in cells. Giant unilamellar vesicles (GUVs)
provide a promising chassis to control, mimic, and understand the
LLPS process; however, they are challenging to construct. Here, we
engineer the dynamic assembly and disassembly of LLPS
compartments using complex coacervates as models inside
synthetic cells. Semipermeable GUVs constructed with defined
lipid composition encapsulate the biomolecules, including enzymes
required to regulate coacervates. Assembly and disassembly of
coacervates are triggered in independent systems by the diffusion
of substrates through the membrane into the vesicle lumen. The
coupling of enzyme networks in a single synthetic cell system allows for reversible and out-of-equilibrium regulation of coacervates.
The functional properties of the coacervates are revealed by sequestering biomolecules, including drugs and enzymes. GUVs, with
functional LLPS compartment assembly, open avenues in constructing programmable autonomous synthetic cells with membraneless
organelles. The coacervate-in-vesicle platform has significant implications for understanding LLPS regulation mechanisms in
cells.
