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Comparison between Lipofectamine RNAiMAX and GenMute transfection agents in two cellular models of human hepatoma

RNA interference is a powerful approach to understand gene function both for therapeutic and experimental purposes. Since the lack of knowledge in the gene silencing of various hepatic cell lines, this work was aimed to compare two transfection agents, the liposome-based Lipofectamine™ RNAiMAX and t...

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Autores principales: Berardo, Clarissa, Siciliano, Veronica, Pasqua, Laura G. Di, Richelmi, Plinio, Vairetti, Mariapia, Ferrigno, Andrea
Formato: Online Artículo Texto
Lenguaje:English
Publicado: PAGEPress Publications, Pavia, Italy 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6712361/
https://www.ncbi.nlm.nih.gov/pubmed/31455073
http://dx.doi.org/10.4081/ejh.2019.3048
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author Berardo, Clarissa
Siciliano, Veronica
Pasqua, Laura G. Di
Richelmi, Plinio
Vairetti, Mariapia
Ferrigno, Andrea
author_facet Berardo, Clarissa
Siciliano, Veronica
Pasqua, Laura G. Di
Richelmi, Plinio
Vairetti, Mariapia
Ferrigno, Andrea
author_sort Berardo, Clarissa
collection PubMed
description RNA interference is a powerful approach to understand gene function both for therapeutic and experimental purposes. Since the lack of knowledge in the gene silencing of various hepatic cell lines, this work was aimed to compare two transfection agents, the liposome-based Lipofectamine™ RNAiMAX and the HepG2-specific, polymer-based GenMute™, in two cellular models of human hepatoma, HepG2 and Huh7.5. In the first part, we assessed transfection efficiency of a fluorescent Cy3-labeled negative control siRNA by cell imaging analysis; we found that cells treated with GenMute present a higher uptake of the fluorescent negative control siRNA when compared to Lipofectamine RNAiMAX-trasfected cells, both in HepG2 and in Huh7.5 cells. In the second part, we evaluated GAPDH silencing with the two transfection reagents by RT-PCR finding similar GAPDH mRNA expression after each transfection treatment. Finally, we measured cell viability by the MTT assay, observing that cells transfected with GenMute have a higher viability with respect to Lipofectamine RNAiMAXadministered cells. These results suggest that GenMute reagent might be considered the most suitable transfection agent for hepatic gene silencing.
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spelling pubmed-67123612019-09-12 Comparison between Lipofectamine RNAiMAX and GenMute transfection agents in two cellular models of human hepatoma Berardo, Clarissa Siciliano, Veronica Pasqua, Laura G. Di Richelmi, Plinio Vairetti, Mariapia Ferrigno, Andrea Eur J Histochem Technical Note RNA interference is a powerful approach to understand gene function both for therapeutic and experimental purposes. Since the lack of knowledge in the gene silencing of various hepatic cell lines, this work was aimed to compare two transfection agents, the liposome-based Lipofectamine™ RNAiMAX and the HepG2-specific, polymer-based GenMute™, in two cellular models of human hepatoma, HepG2 and Huh7.5. In the first part, we assessed transfection efficiency of a fluorescent Cy3-labeled negative control siRNA by cell imaging analysis; we found that cells treated with GenMute present a higher uptake of the fluorescent negative control siRNA when compared to Lipofectamine RNAiMAX-trasfected cells, both in HepG2 and in Huh7.5 cells. In the second part, we evaluated GAPDH silencing with the two transfection reagents by RT-PCR finding similar GAPDH mRNA expression after each transfection treatment. Finally, we measured cell viability by the MTT assay, observing that cells transfected with GenMute have a higher viability with respect to Lipofectamine RNAiMAXadministered cells. These results suggest that GenMute reagent might be considered the most suitable transfection agent for hepatic gene silencing. PAGEPress Publications, Pavia, Italy 2019-08-06 /pmc/articles/PMC6712361/ /pubmed/31455073 http://dx.doi.org/10.4081/ejh.2019.3048 Text en ©Copyright: the Author(s), 2019 http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution Noncommercial License (by-nc 4.0) which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.
spellingShingle Technical Note
Berardo, Clarissa
Siciliano, Veronica
Pasqua, Laura G. Di
Richelmi, Plinio
Vairetti, Mariapia
Ferrigno, Andrea
Comparison between Lipofectamine RNAiMAX and GenMute transfection agents in two cellular models of human hepatoma
title Comparison between Lipofectamine RNAiMAX and GenMute transfection agents in two cellular models of human hepatoma
title_full Comparison between Lipofectamine RNAiMAX and GenMute transfection agents in two cellular models of human hepatoma
title_fullStr Comparison between Lipofectamine RNAiMAX and GenMute transfection agents in two cellular models of human hepatoma
title_full_unstemmed Comparison between Lipofectamine RNAiMAX and GenMute transfection agents in two cellular models of human hepatoma
title_short Comparison between Lipofectamine RNAiMAX and GenMute transfection agents in two cellular models of human hepatoma
title_sort comparison between lipofectamine rnaimax and genmute transfection agents in two cellular models of human hepatoma
topic Technical Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6712361/
https://www.ncbi.nlm.nih.gov/pubmed/31455073
http://dx.doi.org/10.4081/ejh.2019.3048
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