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Formation of phase separated vesicles by double layer cDICE

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Abstract

Recently, continuous droplet interface crossing encapsulation (cDICE) was developed, which allows fast and efficient production of giant unilamellar vesicles (GUVs) under high salt conditions, at low temperature and with low consumption of the encapsulated proteins. Unfortunately, cholesterol encapsulation within the lipid bilayer was not efficient for the cDICE protocol so far and thus the formation of phase separated vesicles was limited. Here we present a modified version of cDICE that allows incorporation of cholesterol into lipid bilayers and enables the reproducible formation of phase-separated vesicles. We show that cholesterol incorporation relies on the amount of mineral oil in the lipid–oil emulsions, which is essential for protein encapsulation inside GUVs by cDICE. The possibility of creating phase separated vesicles by cDICE will enable the study of the interdependence between phase separation and cytoskeletal proteins under confinement.

Graphical abstract: Formation of phase separated vesicles by double layer cDICE

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Publication details

The article was received on 22 Mar 2019, accepted on 16 Jul 2019 and first published on 30 Oct 2019


Article type: Paper
DOI: 10.1039/C8SM02491J
Soft Matter, 2019, Advance Article
  • Open access: Creative Commons BY-NC license
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    Formation of phase separated vesicles by double layer cDICE

    K. Dürre and A. R. Bausch, Soft Matter, 2019, Advance Article , DOI: 10.1039/C8SM02491J

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