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The Prodigious Potential of mRNA Electrotransfer as a Substitute to Conventional DNA-Based Transient Transfection

Transient transfection of foreign DNA is the most widely used laboratory technique to study gene function and product. However, the transfection efficiency depends on many parameters, including DNA quantity and quality, transfection methods and target cell lines. Here, we describe the considerable a...

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Detalles Bibliográficos
Autores principales: Juncker, Théo, Chatton, Bruno, Donzeau, Mariel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10297312/
https://www.ncbi.nlm.nih.gov/pubmed/37371061
http://dx.doi.org/10.3390/cells12121591
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author Juncker, Théo
Chatton, Bruno
Donzeau, Mariel
author_facet Juncker, Théo
Chatton, Bruno
Donzeau, Mariel
author_sort Juncker, Théo
collection PubMed
description Transient transfection of foreign DNA is the most widely used laboratory technique to study gene function and product. However, the transfection efficiency depends on many parameters, including DNA quantity and quality, transfection methods and target cell lines. Here, we describe the considerable advantage of mRNA electroporation compared to conventional DNA-based systems. Indeed, our methodology offers extremely high transfection efficiency up to 98% regardless of the cell line tested. Protein expression takes place a few hours post-transfection and lasts over 72 h, but overall, the electrotransfer of mRNAs enables the monitoring of the level of protein expressed by simply modulating the amount of mRNAs used. As a result, we successfully conducted cell imaging by matching the levels of expressed V(H)Hs and the antigen present in the cell, preventing the necessity to remove the excess unbound V(H)Hs. Altogether, our results demonstrate that mRNA electrotransfer could easily supplant the conventional DNA-based transient expression system.
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spelling pubmed-102973122023-06-28 The Prodigious Potential of mRNA Electrotransfer as a Substitute to Conventional DNA-Based Transient Transfection Juncker, Théo Chatton, Bruno Donzeau, Mariel Cells Communication Transient transfection of foreign DNA is the most widely used laboratory technique to study gene function and product. However, the transfection efficiency depends on many parameters, including DNA quantity and quality, transfection methods and target cell lines. Here, we describe the considerable advantage of mRNA electroporation compared to conventional DNA-based systems. Indeed, our methodology offers extremely high transfection efficiency up to 98% regardless of the cell line tested. Protein expression takes place a few hours post-transfection and lasts over 72 h, but overall, the electrotransfer of mRNAs enables the monitoring of the level of protein expressed by simply modulating the amount of mRNAs used. As a result, we successfully conducted cell imaging by matching the levels of expressed V(H)Hs and the antigen present in the cell, preventing the necessity to remove the excess unbound V(H)Hs. Altogether, our results demonstrate that mRNA electrotransfer could easily supplant the conventional DNA-based transient expression system. MDPI 2023-06-08 /pmc/articles/PMC10297312/ /pubmed/37371061 http://dx.doi.org/10.3390/cells12121591 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 Communication
Juncker, Théo
Chatton, Bruno
Donzeau, Mariel
The Prodigious Potential of mRNA Electrotransfer as a Substitute to Conventional DNA-Based Transient Transfection
title The Prodigious Potential of mRNA Electrotransfer as a Substitute to Conventional DNA-Based Transient Transfection
title_full The Prodigious Potential of mRNA Electrotransfer as a Substitute to Conventional DNA-Based Transient Transfection
title_fullStr The Prodigious Potential of mRNA Electrotransfer as a Substitute to Conventional DNA-Based Transient Transfection
title_full_unstemmed The Prodigious Potential of mRNA Electrotransfer as a Substitute to Conventional DNA-Based Transient Transfection
title_short The Prodigious Potential of mRNA Electrotransfer as a Substitute to Conventional DNA-Based Transient Transfection
title_sort prodigious potential of mrna electrotransfer as a substitute to conventional dna-based transient transfection
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10297312/
https://www.ncbi.nlm.nih.gov/pubmed/37371061
http://dx.doi.org/10.3390/cells12121591
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