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Rapid and Facile Preparation of Giant Vesicles by the Droplet Transfer Method for Artificial Cell Construction
Giant vesicles have been widely used for the bottom-up construction of artificial (or synthetic) cells and the physicochemical analysis of lipid membranes. Although methods for the formation of giant vesicles and the encapsulation of molecules within them have been established, a standardized protoc...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2022
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9021372/ https://www.ncbi.nlm.nih.gov/pubmed/35464723 http://dx.doi.org/10.3389/fbioe.2022.873854 |
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author | Shimane, Yasuhiro Kuruma, Yutetsu |
author_facet | Shimane, Yasuhiro Kuruma, Yutetsu |
author_sort | Shimane, Yasuhiro |
collection | PubMed |
description | Giant vesicles have been widely used for the bottom-up construction of artificial (or synthetic) cells and the physicochemical analysis of lipid membranes. Although methods for the formation of giant vesicles and the encapsulation of molecules within them have been established, a standardized protocol has not been shared among researchers including non-experts. Here we proposed a rapid and facile protocol that allows the formation of giant vesicles within 30 min. The quality of the giant vesicles encapsulating a cell-free protein expression system was comparable to that of the ones formed using a conventional method, in terms of the synthesis of both soluble and membrane proteins. We also performed protein synthesis in artificial cells using a lyophilized cell-free mixture and showed an equivalent level of protein synthesis. Our method could become a standard method for giant vesicle formation suited for artificial cell research. |
format | Online Article Text |
id | pubmed-9021372 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-90213722022-04-22 Rapid and Facile Preparation of Giant Vesicles by the Droplet Transfer Method for Artificial Cell Construction Shimane, Yasuhiro Kuruma, Yutetsu Front Bioeng Biotechnol Bioengineering and Biotechnology Giant vesicles have been widely used for the bottom-up construction of artificial (or synthetic) cells and the physicochemical analysis of lipid membranes. Although methods for the formation of giant vesicles and the encapsulation of molecules within them have been established, a standardized protocol has not been shared among researchers including non-experts. Here we proposed a rapid and facile protocol that allows the formation of giant vesicles within 30 min. The quality of the giant vesicles encapsulating a cell-free protein expression system was comparable to that of the ones formed using a conventional method, in terms of the synthesis of both soluble and membrane proteins. We also performed protein synthesis in artificial cells using a lyophilized cell-free mixture and showed an equivalent level of protein synthesis. Our method could become a standard method for giant vesicle formation suited for artificial cell research. Frontiers Media S.A. 2022-04-07 /pmc/articles/PMC9021372/ /pubmed/35464723 http://dx.doi.org/10.3389/fbioe.2022.873854 Text en Copyright © 2022 Shimane and Kuruma. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Bioengineering and Biotechnology Shimane, Yasuhiro Kuruma, Yutetsu Rapid and Facile Preparation of Giant Vesicles by the Droplet Transfer Method for Artificial Cell Construction |
title | Rapid and Facile Preparation of Giant Vesicles by the Droplet Transfer Method for Artificial Cell Construction |
title_full | Rapid and Facile Preparation of Giant Vesicles by the Droplet Transfer Method for Artificial Cell Construction |
title_fullStr | Rapid and Facile Preparation of Giant Vesicles by the Droplet Transfer Method for Artificial Cell Construction |
title_full_unstemmed | Rapid and Facile Preparation of Giant Vesicles by the Droplet Transfer Method for Artificial Cell Construction |
title_short | Rapid and Facile Preparation of Giant Vesicles by the Droplet Transfer Method for Artificial Cell Construction |
title_sort | rapid and facile preparation of giant vesicles by the droplet transfer method for artificial cell construction |
topic | Bioengineering and Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9021372/ https://www.ncbi.nlm.nih.gov/pubmed/35464723 http://dx.doi.org/10.3389/fbioe.2022.873854 |
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