Spatiotemporal control of coacervate formation within liposomes

Nat Commun. 2019 Apr 17;10(1):1800. doi: 10.1038/s41467-019-09855-x.

Abstract

Liquid-liquid phase separation (LLPS), especially coacervation, plays a crucial role in cell biology, as it forms numerous membraneless organelles in cells. Coacervates play an indispensable role in regulating intracellular biochemistry, and their dysfunction is associated with several diseases. Understanding of the LLPS dynamics would greatly benefit from controlled in vitro assays that mimic cells. Here, we use a microfluidics-based methodology to form coacervates inside cell-sized (~10 µm) liposomes, allowing control over the dynamics. Protein-pore-mediated permeation of small molecules into liposomes triggers LLPS passively or via active mechanisms like enzymatic polymerization of nucleic acids. We demonstrate sequestration of proteins (FtsZ) and supramolecular assemblies (lipid vesicles), as well as the possibility to host metabolic reactions (β-galactosidase activity) inside coacervates. This coacervate-in-liposome platform provides a versatile tool to understand intracellular phase behavior, and these hybrid systems will allow engineering complex pathways to reconstitute cellular functions and facilitate bottom-up creation of synthetic cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Artificial Cells / chemistry
  • Artificial Cells / metabolism*
  • Cell Membrane Permeability*
  • High-Throughput Screening Assays / instrumentation
  • High-Throughput Screening Assays / methods
  • Lab-On-A-Chip Devices
  • Liposomes / chemistry
  • Liposomes / metabolism*
  • Microfluidics / instrumentation
  • Microfluidics / methods*
  • Phase Transition

Substances

  • Liposomes