Fiche publication
Date publication
juin 2023
Journal
Cells
Auteurs
Membres identifiés du Cancéropôle Est :
Pr CHATTON Bruno
,
Dr DONZEAU Mariel
Tous les auteurs :
Juncker T, Chatton B, Donzeau M
Lien Pubmed
Résumé
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 VHs and the antigen present in the cell, preventing the necessity to remove the excess unbound VHs. Altogether, our results demonstrate that mRNA electrotransfer could easily supplant the conventional DNA-based transient expression system.
Mots clés
cell imaging, electroporation, mRNAs, nanobody, transfection efficiency, transient transfection
Référence
Cells. 2023 06 8;12(12):
