Sequential bottom-up assembly of mechanically stabilized synthetic cells by microfluidics
Top Cited Papers
- 16 October 2017
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Materials
- Vol. 17 (1), 89-96
- https://doi.org/10.1038/nmat5005
Abstract
Compartments for the spatially and temporally controlled assembly of biological processes are essential towards cellular life. Synthetic mimics of cellular compartments based on lipid-based protocells lack the mechanical and chemical stability to allow their manipulation into a complex and fully functional synthetic cell. Here, we present a high-throughput microfluidic method to generate stable, defined sized liposomes termed ‘droplet-stabilized giant unilamellar vesicles (dsGUVs)’. The enhanced stability of dsGUVs enables the sequential loading of these compartments with biomolecules, namely purified transmembrane and cytoskeleton proteins by microfluidic pico-injection technology. This constitutes an experimental demonstration of a successful bottom-up assembly of a compartment with contents that would not self-assemble to full functionality when simply mixed together. Following assembly, the stabilizing oil phase and droplet shells are removed to release functional self-supporting protocells to an aqueous phase, enabling them to interact with physiologically relevant matrices.This publication has 49 references indexed in Scilit:
- Synthesis of Nanostructured and Biofunctionalized Water-in-Oil Droplets as Tools for Homing T CellsJournal of the American Chemical Society, 2013
- Polymer Vesicles as Microreactors for Bioinspired Calcium Carbonate PrecipitationLangmuir, 2011
- Stepwise Synthesis of Giant Unilamellar Vesicles on a Microfluidic Assembly LineJournal of the American Chemical Society, 2011
- High-throughput injection with microfluidics using picoinjectorsProceedings of the National Academy of Sciences of the United States of America, 2010
- Lipid diffusion in planar membranes investigated by fluorescence correlation spectroscopyBiochimica et Biophysica Acta (BBA) - Biomembranes, 2010
- Integrin reconstituted in GUVs: A biomimetic system to study initial steps of cell spreadingBiochimica et Biophysica Acta (BBA) - Biomembranes, 2009
- Environmental sensing through focal adhesionsNature Reviews Molecular Cell Biology, 2009
- Functionalized nanocompartments (Synthosomes): Limitations and prospective applications in industrial biotechnologyBiotechnology Journal, 2006
- Purification of brain tubulin through two cycles of polymerization–depolymerization in a high-molarity bufferProtein Expression and Purification, 2003
- Comparison of ΔpH‐ and Δφ‐driven ATP synthesis catalyzed by the H+‐ATPases from Escherichia coli or chloroplasts reconstituted into liposomesFEBS Letters, 1999