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Construction of Fusion Protein for Enhanced Small RNA Loading to Extracellular Vesicles
Extracellular vesicles (EVs) naturally carry cargo from producer cells, such as RNA and protein, and can transfer these messengers to other cells and tissue. This ability provides an interesting opportunity for using EVs as delivery vehicles for therapeutic agents, such as for gene therapy. However,...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9956110/ https://www.ncbi.nlm.nih.gov/pubmed/36833188 http://dx.doi.org/10.3390/genes14020261 |
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author | Es-Haghi, Masoumeh Neustroeva, Olga Chowdhury, Iftekhar Laitinen, Pia Väänänen, Mari-Anna Korvenlaita, Nea Malm, Tarja Turunen, Mikko P. Turunen, Tiia A. |
author_facet | Es-Haghi, Masoumeh Neustroeva, Olga Chowdhury, Iftekhar Laitinen, Pia Väänänen, Mari-Anna Korvenlaita, Nea Malm, Tarja Turunen, Mikko P. Turunen, Tiia A. |
author_sort | Es-Haghi, Masoumeh |
collection | PubMed |
description | Extracellular vesicles (EVs) naturally carry cargo from producer cells, such as RNA and protein, and can transfer these messengers to other cells and tissue. This ability provides an interesting opportunity for using EVs as delivery vehicles for therapeutic agents, such as for gene therapy. However, endogenous loading of cargo, such as microRNAs (miRNAs), is not very efficient as the copy number of miRNAs per EV is quite low. Therefore, new methods and tools to enhance the loading of small RNAs is required. In the current study, we developed fusion protein of EV membrane protein CD9 and RNA-binding protein AGO2 (hCD9.hAGO2). We show that the EVs engineered with hCD9.hAGO2 contain significantly higher levels of miRNA or shRNA (miR-466c or shRNA-451, respectively) compared to EVs that are isolated from cells that only overexpress the desired miRNA or shRNA. These hCD9.hAGO2 engineered EVs also transfer their RNA cargo to recipient cells more efficiently. We were not able to detect changes in gene expression levels in recipient cells after the EV treatments, but we show that the cell viability of HUVECs was increased after hCD9.hAGO2 EV treatments. This technical study characterizes the hCD9.hAGO2 fusion protein for the future development of enhanced RNA loading to EVs. |
format | Online Article Text |
id | pubmed-9956110 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-99561102023-02-25 Construction of Fusion Protein for Enhanced Small RNA Loading to Extracellular Vesicles Es-Haghi, Masoumeh Neustroeva, Olga Chowdhury, Iftekhar Laitinen, Pia Väänänen, Mari-Anna Korvenlaita, Nea Malm, Tarja Turunen, Mikko P. Turunen, Tiia A. Genes (Basel) Article Extracellular vesicles (EVs) naturally carry cargo from producer cells, such as RNA and protein, and can transfer these messengers to other cells and tissue. This ability provides an interesting opportunity for using EVs as delivery vehicles for therapeutic agents, such as for gene therapy. However, endogenous loading of cargo, such as microRNAs (miRNAs), is not very efficient as the copy number of miRNAs per EV is quite low. Therefore, new methods and tools to enhance the loading of small RNAs is required. In the current study, we developed fusion protein of EV membrane protein CD9 and RNA-binding protein AGO2 (hCD9.hAGO2). We show that the EVs engineered with hCD9.hAGO2 contain significantly higher levels of miRNA or shRNA (miR-466c or shRNA-451, respectively) compared to EVs that are isolated from cells that only overexpress the desired miRNA or shRNA. These hCD9.hAGO2 engineered EVs also transfer their RNA cargo to recipient cells more efficiently. We were not able to detect changes in gene expression levels in recipient cells after the EV treatments, but we show that the cell viability of HUVECs was increased after hCD9.hAGO2 EV treatments. This technical study characterizes the hCD9.hAGO2 fusion protein for the future development of enhanced RNA loading to EVs. MDPI 2023-01-19 /pmc/articles/PMC9956110/ /pubmed/36833188 http://dx.doi.org/10.3390/genes14020261 Text en © 2023 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 | Article Es-Haghi, Masoumeh Neustroeva, Olga Chowdhury, Iftekhar Laitinen, Pia Väänänen, Mari-Anna Korvenlaita, Nea Malm, Tarja Turunen, Mikko P. Turunen, Tiia A. Construction of Fusion Protein for Enhanced Small RNA Loading to Extracellular Vesicles |
title | Construction of Fusion Protein for Enhanced Small RNA Loading to Extracellular Vesicles |
title_full | Construction of Fusion Protein for Enhanced Small RNA Loading to Extracellular Vesicles |
title_fullStr | Construction of Fusion Protein for Enhanced Small RNA Loading to Extracellular Vesicles |
title_full_unstemmed | Construction of Fusion Protein for Enhanced Small RNA Loading to Extracellular Vesicles |
title_short | Construction of Fusion Protein for Enhanced Small RNA Loading to Extracellular Vesicles |
title_sort | construction of fusion protein for enhanced small rna loading to extracellular vesicles |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9956110/ https://www.ncbi.nlm.nih.gov/pubmed/36833188 http://dx.doi.org/10.3390/genes14020261 |
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