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Sepiolite as a new nanocarrier for DNA transfer into mammalian cells

Jeudi 4 octobre 2018 11:30 - Duree : 1 heure
Lieu : CEA, Bâtiment C5, Salle 421A - 17 rue des martyrs - 38000 Grenoble

Orateur : Bernard S. LOPEZ (Institut Gustave Roussy, Villejuif)

Abstract :

Nanofibers of sepiolite, a natural silicate belonging to the clay minerals family, might constitute a potential promising nanocarrier for the non-viral transfer of bio-molecules. Using physic-chemical approach and transmission electronic analyses, we show that sepiolite nanofibers efficiently bind different types of DNA molecules through electrostatic interactions, hydrogen bonding, cation bridges, and van der Waals forces. Moreover, Fourier-transform infrared spectroscopy identified the external silanol groups as the main sites of interaction with the DNA.

Due to its nano-size dimension sepiolite can be naturally internalized into mammalian cells. Therefore, deciphering the mechanisms of sepiolite cell internalization constitutes a question interesting biotechnology, for the use of sepiolite as nanocarrier, as well as environmental and public health concerns. Though it is low, the perfectly stable and natural intrinsic fluorescence of sepiolite nanofibers allows to follow their fate into cells. By combining fluorescence microscopy, time-lapse video microscopy, fluorescence activated cell sorting and transmission electron microscopy, we show that sepiolite can be spontaneously internalized into mammalian cells through both non-endocytic and endocytic pathways, macropinocytosis being one of the main pathways.

As a proof of concept, we show that sepiolite is able to stably transfer plasmid DNA into mammalian cells and that the efficiency can be optimized. Indeed, sonication of sepiolite prior assembly with DNA 100-fold stimulated DNA transfection efficiency. Moreover, exposure of the cells to endocytosis inhibitors, such as chloroquine, two-fold increase the efficiency of sepiolite-mediated gene transfer, in addition to the 100-fold increased resulting from sepiolite sonomechanical treatment.

These results open the way to the use of sepiolite-based bionanocomposites as a novel class of nanoplatform for gene transfer.

Contact : Jean-Luc Ravanat - 04.38.78.47.97

Discipline évènement : (Biologie / Chimie)
Entité organisatrice : (INAC/SyMMES)
Nature évènement : (Séminaire)
Site de l'évènement : Site CEA avec accès badge

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