Regulating Biocondensates within Synthetic Cells via Segregative Phase Separation

Area of Science:

• Biophysics
• Cell Biology
• Synthetic Biology

Background:

• Cells utilize macromolecular interactions to drive phase separation, creating dynamic intracellular compartments.
• The influence of simultaneous, multiple phase separation events on cellular organization remains largely unexplored.

Purpose of the Study:

• To investigate the interplay between segregative and associative phase separation within cell-mimicking confinements.
• To understand how these phenomena influence intracellular compartmentalization and lipid boundary interactions.

Main Methods:

• Utilized on-chip microfluidic systems to encapsulate associative and segregative components.
• Externally triggered phase separation within cell-sized vesicles to mimic cellular environments.

Main Results:

• Segregative phases formed microdomains, influencing associative components as recruiters, membrane-targeting agents, and condensation initiators.
• Multiphase architecture created isolated microenvironments, restricting molecular communication and diffusion.
• Observed global shape transformation of the confinement due to hierarchical domain formation at the lipid membrane.

Conclusions:

• Segregative phase separation is proposed as a universal strategy for regulating condensate formation and localization.
• The interplay between phase separation systems provides design principles for synthetic cells and artificial membraneless organelles.