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Dynamic Control of Functional Coacervates in Synthetic Cells.

Karthika S Nair1,2, Sreelakshmi Radhakrishnan1, Harsha Bajaj1,2

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Scientists engineered dynamic, membrane-less compartments using complex coacervates within synthetic cells. This breakthrough enables controlled assembly and disassembly of liquid-liquid phase separation (LLPS) compartments, advancing synthetic biology.

Keywords:
compartmentalizationcomplex coacervatesgiant unilamellar vesiclesliquid−liquid phase separationout-of-equilibrium systemsynthetic cells

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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.