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Use of Bacterial Extracellular Vesicles for Gene Delivery to Host Cells

Extracellular vesicles (EVs), which are nanosized membranous particles secreted from both prokaryotic and eukaryotic cells, can deliver various biological molecules, such as nucleic acids, proteins, and lipids, into recipient cells. However, contrary to what is known about eukaryotic EVs, whether ba...

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Autores principales: Kim, Su-Im, Ha, Jae Yeong, Choi, Song-Yi, Hong, Su-Hyung, Lee, Heon-Jin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9496234/
https://www.ncbi.nlm.nih.gov/pubmed/36139009
http://dx.doi.org/10.3390/biom12091171
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author Kim, Su-Im
Ha, Jae Yeong
Choi, Song-Yi
Hong, Su-Hyung
Lee, Heon-Jin
author_facet Kim, Su-Im
Ha, Jae Yeong
Choi, Song-Yi
Hong, Su-Hyung
Lee, Heon-Jin
author_sort Kim, Su-Im
collection PubMed
description Extracellular vesicles (EVs), which are nanosized membranous particles secreted from both prokaryotic and eukaryotic cells, can deliver various biological molecules, such as nucleic acids, proteins, and lipids, into recipient cells. However, contrary to what is known about eukaryotic EVs, whether bacterial EVs (bEVs) can be used as transporters for bioactive molecules is becoming a hot area of research. In this study, we electroporated enhanced green fluorescent protein (EGFP) genes and precursor microRNA of Cel-miR-39 (pre-Cel-miR-39) from isolated bEVs of Escherichia coli and Lactobacillus reuteri. The EGFP plasmid, synthetic EGFP RNA, and pre-Cel-miR-39 were successfully delivered into the murine microglial BV2 cells via bEVs. PCR and confocal microscopy analysis confirmed the transfer of the EGFP plasmid and RNA. The bEV-delivered exogenous pre-Cel-miR-39 was further processed into the mature form of Cel-miR-39; its incorporation into Ago2—a major component of the RNA-induced silencing complex—was assessed using RNA-immunoprecipitation–PCR. Taken together, bEVs can be used as vehicles to deliver genetic materials and for novel biotechnological applications, such as gene transfer and mRNA vaccines.
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spelling pubmed-94962342022-09-23 Use of Bacterial Extracellular Vesicles for Gene Delivery to Host Cells Kim, Su-Im Ha, Jae Yeong Choi, Song-Yi Hong, Su-Hyung Lee, Heon-Jin Biomolecules Communication Extracellular vesicles (EVs), which are nanosized membranous particles secreted from both prokaryotic and eukaryotic cells, can deliver various biological molecules, such as nucleic acids, proteins, and lipids, into recipient cells. However, contrary to what is known about eukaryotic EVs, whether bacterial EVs (bEVs) can be used as transporters for bioactive molecules is becoming a hot area of research. In this study, we electroporated enhanced green fluorescent protein (EGFP) genes and precursor microRNA of Cel-miR-39 (pre-Cel-miR-39) from isolated bEVs of Escherichia coli and Lactobacillus reuteri. The EGFP plasmid, synthetic EGFP RNA, and pre-Cel-miR-39 were successfully delivered into the murine microglial BV2 cells via bEVs. PCR and confocal microscopy analysis confirmed the transfer of the EGFP plasmid and RNA. The bEV-delivered exogenous pre-Cel-miR-39 was further processed into the mature form of Cel-miR-39; its incorporation into Ago2—a major component of the RNA-induced silencing complex—was assessed using RNA-immunoprecipitation–PCR. Taken together, bEVs can be used as vehicles to deliver genetic materials and for novel biotechnological applications, such as gene transfer and mRNA vaccines. MDPI 2022-08-24 /pmc/articles/PMC9496234/ /pubmed/36139009 http://dx.doi.org/10.3390/biom12091171 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Kim, Su-Im
Ha, Jae Yeong
Choi, Song-Yi
Hong, Su-Hyung
Lee, Heon-Jin
Use of Bacterial Extracellular Vesicles for Gene Delivery to Host Cells
title Use of Bacterial Extracellular Vesicles for Gene Delivery to Host Cells
title_full Use of Bacterial Extracellular Vesicles for Gene Delivery to Host Cells
title_fullStr Use of Bacterial Extracellular Vesicles for Gene Delivery to Host Cells
title_full_unstemmed Use of Bacterial Extracellular Vesicles for Gene Delivery to Host Cells
title_short Use of Bacterial Extracellular Vesicles for Gene Delivery to Host Cells
title_sort use of bacterial extracellular vesicles for gene delivery to host cells
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9496234/
https://www.ncbi.nlm.nih.gov/pubmed/36139009
http://dx.doi.org/10.3390/biom12091171
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