pH-Triggered Assembly of Endomembrane Multicompartments in Synthetic Cells

Area of Science:

• Synthetic Biology
• Biophysics
• Materials Science

Background:

• Droplet-stabilized giant unilamellar vesicles (GUVs) are used to create minimal synthetic cells.
• Current methods face low production efficiency due to difficulties in releasing GUVs from stabilizing droplets.

Purpose of the Study:

• To develop a pH-triggered method for efficient self-assembly and release of droplet-stabilized GUVs (dsGUVs).
• To enhance the production efficiency and robustness of compartmentalized GUVs for synthetic cell applications.

Main Methods:

• Utilized pH as a trigger for self-assembly of dsGUVs via bulk or droplet-based microfluidics.
• Co-encapsulated pH-sensitive small unilamellar vesicles (SUVs) and/or proteins within the GUVs.
• Investigated the effect of droplet acidification on GUV formation and sequestration of encapsulated materials.

Main Results:

• Demonstrated efficient production of dsGUVs upon droplet acidification.
• Showcased the ability to sequester co-encapsulated materials, including pH-sensitive SUVs and proteins.
• Significantly improved the production efficiency of free-standing GUVs compared to previous methods.

Conclusions:

• pH-mediated assembly offers a flexible and robust strategy for generating compartmentalized dsGUVs.
• This approach overcomes major production hurdles, paving the way for more efficient synthetic cell development.
• The pH-triggered method enhances GUV release, advancing the field of minimal synthetic cells.