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Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method

Messenger RNA (mRNA)-based therapies are a novel class of therapeutics used in vaccination and protein replacement therapies for monogenic diseases. Previously, we developed a modified ethanol injection (MEI) method for small interfering RNA (siRNA) transfection, in which cationic liposome/siRNA com...

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Autores principales: Tang, Min, Sagawa, Ayane, Inoue, Nodoka, Torii, Satomi, Tomita, Kana, Hattori, Yoshiyuki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10146866/
https://www.ncbi.nlm.nih.gov/pubmed/37111627
http://dx.doi.org/10.3390/pharmaceutics15041141
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author Tang, Min
Sagawa, Ayane
Inoue, Nodoka
Torii, Satomi
Tomita, Kana
Hattori, Yoshiyuki
author_facet Tang, Min
Sagawa, Ayane
Inoue, Nodoka
Torii, Satomi
Tomita, Kana
Hattori, Yoshiyuki
author_sort Tang, Min
collection PubMed
description Messenger RNA (mRNA)-based therapies are a novel class of therapeutics used in vaccination and protein replacement therapies for monogenic diseases. Previously, we developed a modified ethanol injection (MEI) method for small interfering RNA (siRNA) transfection, in which cationic liposome/siRNA complexes (siRNA lipoplexes) were prepared by mixing a lipid-ethanol solution with a siRNA solution. In this study, we applied the MEI method to prepare mRNA lipoplexes and evaluated the in vitro and in vivo protein expression efficiencies. We selected six cationic lipids and three neutral helper lipids to generate 18 mRNA lipoplexes. These were composed of cationic lipids, neutral helper lipids, and polyethylene glycol-cholesteryl ether (PEG-Chol). Among them, mRNA lipoplexes containing N-hexadecyl-N,N-dimethylhexadecan-1-aminium bromide (DC-1-16) or 11-((1,3-bis(dodecanoyloxy)-2-((dodecanoyloxy)methyl) propan-2-yl) amino)-N,N,N-trimethyl-11-oxoundecan-1-aminium bromide (TC-1-12) with 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and PEG-Chol exhibited high protein expression in cells. Furthermore, mRNA lipoplexes composed of DC-1-16, DOPE, and PEG-Chol exhibited high protein expression in the lungs and spleen of mice after systemic injection and induced high antigen-specific IgG1 levels upon immunization. These results suggest that the MEI method can potentially increase the efficiency of mRNA transfection, both in vitro and in vivo.
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spelling pubmed-101468662023-04-29 Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method Tang, Min Sagawa, Ayane Inoue, Nodoka Torii, Satomi Tomita, Kana Hattori, Yoshiyuki Pharmaceutics Article Messenger RNA (mRNA)-based therapies are a novel class of therapeutics used in vaccination and protein replacement therapies for monogenic diseases. Previously, we developed a modified ethanol injection (MEI) method for small interfering RNA (siRNA) transfection, in which cationic liposome/siRNA complexes (siRNA lipoplexes) were prepared by mixing a lipid-ethanol solution with a siRNA solution. In this study, we applied the MEI method to prepare mRNA lipoplexes and evaluated the in vitro and in vivo protein expression efficiencies. We selected six cationic lipids and three neutral helper lipids to generate 18 mRNA lipoplexes. These were composed of cationic lipids, neutral helper lipids, and polyethylene glycol-cholesteryl ether (PEG-Chol). Among them, mRNA lipoplexes containing N-hexadecyl-N,N-dimethylhexadecan-1-aminium bromide (DC-1-16) or 11-((1,3-bis(dodecanoyloxy)-2-((dodecanoyloxy)methyl) propan-2-yl) amino)-N,N,N-trimethyl-11-oxoundecan-1-aminium bromide (TC-1-12) with 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and PEG-Chol exhibited high protein expression in cells. Furthermore, mRNA lipoplexes composed of DC-1-16, DOPE, and PEG-Chol exhibited high protein expression in the lungs and spleen of mice after systemic injection and induced high antigen-specific IgG1 levels upon immunization. These results suggest that the MEI method can potentially increase the efficiency of mRNA transfection, both in vitro and in vivo. MDPI 2023-04-04 /pmc/articles/PMC10146866/ /pubmed/37111627 http://dx.doi.org/10.3390/pharmaceutics15041141 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
Tang, Min
Sagawa, Ayane
Inoue, Nodoka
Torii, Satomi
Tomita, Kana
Hattori, Yoshiyuki
Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method
title Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method
title_full Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method
title_fullStr Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method
title_full_unstemmed Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method
title_short Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method
title_sort efficient mrna delivery with mrna lipoplexes prepared using a modified ethanol injection method
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10146866/
https://www.ncbi.nlm.nih.gov/pubmed/37111627
http://dx.doi.org/10.3390/pharmaceutics15041141
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