Dynamic Control of Functional Coacervates in Synthetic Cells

Area of Science:

• Biochemistry
• Synthetic Biology
• Materials Science

Background:

• Cellular processes rely on dynamic, membrane-less compartments formed by liquid-liquid phase separation (LLPS).
• Enzyme reactions regulate these LLPS compartments, but mimicking this control in synthetic systems is challenging.
• Giant unilamellar vesicles (GUVs) offer a platform for studying LLPS but are difficult to create and control.

Purpose of the Study:

• To engineer dynamic assembly and disassembly of LLPS compartments using complex coacervates within synthetic cells.
• To demonstrate reversible, out-of-equilibrium regulation of coacervates via coupled enzyme networks.
• To explore the functional properties of coacervates for applications like biomolecule sequestration.

Main Methods:

• Constructed semipermeable GUVs with defined lipid compositions to encapsulate biomolecules and enzymes.
• Utilized complex coacervates as model systems for LLPS compartments inside GUVs.
• Triggered coacervate assembly/disassembly via substrate diffusion through the GUV membrane, controlling enzyme activity.

Main Results:

• Successfully engineered dynamic LLPS compartments within synthetic cells (GUVs).
• Demonstrated enzyme-mediated, substrate-triggered assembly and disassembly of coacervates.
• Showcased reversible, out-of-equilibrium regulation of LLPS through coupled enzyme networks.
• Revealed functional properties of coacervates, including sequestration of drugs and enzymes.

Conclusions:

• The coacervate-in-vesicle platform enables programmable control over membrane-less organelles.
• This system provides a novel approach to understanding LLPS regulation mechanisms relevant to cellular biology.
• Engineered GUVs with functional LLPS compartments open new possibilities for creating autonomous synthetic cells.