Elastin-like polypeptide coacervates as reversibly triggerable compartments for synthetic cells

Area of Science:

• Synthetic biology
• Biomaterials science
• Microfluidics

Background:

• Cellular compartmentalization is crucial for biological function.
• Synthetic cells require dynamic and responsive sub-compartments for engineered processes.
• Membraneless organelles (MOs) formed via coacervation offer a promising approach for artificial cell engineering.

Purpose of the Study:

• To develop a lab-on-a-chip system for reversible triggering of peptide-based coacervates within synthetic cells.
• To engineer dynamic, responsive membraneless organelles (MOs) in artificial cell constructs.
• To demonstrate the utility of elastin-like polypeptides (ELPs) and double emulsion droplets (DEs) for controlled compartmentalization.

Main Methods:

• High-throughput microfluidic production of double emulsion droplets (DEs) encapsulating elastin-like polypeptides (ELPs).
• Utilized pH and temperature variations to induce coacervate formation and dissolution cycles within DEs.
• Developed an integrated microfluidic device for trapping and environmental stimulation of DEs, including osmolyte-based triggering.

Main Results:

• Successfully produced microfluidic DEs with efficient ELP encapsulation.
• Demonstrated multiple cycles of MO formation and dissolution in DEs using physicochemical triggers.
• Showcased reversible compartmentalization within synthetic cells using ELPs and DEs on a microfluidic platform.

Conclusions:

• Engineered reversible membraneless compartmentalization in synthetic cells using DEs and ELPs.
• The microfluidic platform enables precise control over MO dynamics via environmental stimuli.
• The developed system has broad applicability for phase separation and vesicle systems in synthetic cell research.