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Microfluidic platform enables tailored translocation and reaction cascades in nanoliter droplet networks

In the field of bottom-up synthetic biology, lipid membranes are the scaffold to create minimal cells and mimic reactions and processes at or across the membrane. In this context, we employ here a versatile microfluidic platform that enables precise positioning of nanoliter droplets with user-specif...

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Autores principales: Bachler, Simon, Haidas, Dominik, Ort, Marion, Duncombe, Todd A., Dittrich, Petra S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7736871/
https://www.ncbi.nlm.nih.gov/pubmed/33318607
http://dx.doi.org/10.1038/s42003-020-01489-w
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author Bachler, Simon
Haidas, Dominik
Ort, Marion
Duncombe, Todd A.
Dittrich, Petra S.
author_facet Bachler, Simon
Haidas, Dominik
Ort, Marion
Duncombe, Todd A.
Dittrich, Petra S.
author_sort Bachler, Simon
collection PubMed
description In the field of bottom-up synthetic biology, lipid membranes are the scaffold to create minimal cells and mimic reactions and processes at or across the membrane. In this context, we employ here a versatile microfluidic platform that enables precise positioning of nanoliter droplets with user-specified lipid compositions and in a defined pattern. Adjacent droplets make contact and form a droplet interface bilayer to simulate cellular membranes. Translocation of molecules across membranes are tailored by the addition of alpha-hemolysin to selected droplets. Moreover, we developed a protocol to analyze the translocation of non-fluorescent molecules between droplets with mass spectrometry. Our method is capable of automated formation of one- and two-dimensional droplet networks, which we demonstrated by connecting droplets containing different compound and enzyme solutions to perform translocation experiments and a multistep enzymatic cascade reaction across the droplet network. Our platform opens doors for creating complex artificial systems for bottom-up synthetic biology.
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spelling pubmed-77368712020-12-21 Microfluidic platform enables tailored translocation and reaction cascades in nanoliter droplet networks Bachler, Simon Haidas, Dominik Ort, Marion Duncombe, Todd A. Dittrich, Petra S. Commun Biol Article In the field of bottom-up synthetic biology, lipid membranes are the scaffold to create minimal cells and mimic reactions and processes at or across the membrane. In this context, we employ here a versatile microfluidic platform that enables precise positioning of nanoliter droplets with user-specified lipid compositions and in a defined pattern. Adjacent droplets make contact and form a droplet interface bilayer to simulate cellular membranes. Translocation of molecules across membranes are tailored by the addition of alpha-hemolysin to selected droplets. Moreover, we developed a protocol to analyze the translocation of non-fluorescent molecules between droplets with mass spectrometry. Our method is capable of automated formation of one- and two-dimensional droplet networks, which we demonstrated by connecting droplets containing different compound and enzyme solutions to perform translocation experiments and a multistep enzymatic cascade reaction across the droplet network. Our platform opens doors for creating complex artificial systems for bottom-up synthetic biology. Nature Publishing Group UK 2020-12-14 /pmc/articles/PMC7736871/ /pubmed/33318607 http://dx.doi.org/10.1038/s42003-020-01489-w Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Bachler, Simon
Haidas, Dominik
Ort, Marion
Duncombe, Todd A.
Dittrich, Petra S.
Microfluidic platform enables tailored translocation and reaction cascades in nanoliter droplet networks
title Microfluidic platform enables tailored translocation and reaction cascades in nanoliter droplet networks
title_full Microfluidic platform enables tailored translocation and reaction cascades in nanoliter droplet networks
title_fullStr Microfluidic platform enables tailored translocation and reaction cascades in nanoliter droplet networks
title_full_unstemmed Microfluidic platform enables tailored translocation and reaction cascades in nanoliter droplet networks
title_short Microfluidic platform enables tailored translocation and reaction cascades in nanoliter droplet networks
title_sort microfluidic platform enables tailored translocation and reaction cascades in nanoliter droplet networks
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7736871/
https://www.ncbi.nlm.nih.gov/pubmed/33318607
http://dx.doi.org/10.1038/s42003-020-01489-w
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