To date, efficiency upon non-viral DNA delivery remains low and this implies the existence of unidentified transfection barriers. Here we explore the mechanisms of action of multicomponent (MC) cationic liposome/DNA complexes (lipoplexes) by a combination of reporter technologies, dynamic light scattering (DLS), synchrotron small angle X-ray scattering (SAXS), fluorescence activated cell sorting (FACS) analysis and laser scanning confocal microscopy (LSCM) in live cells. Lipofectamine – the gold standard among transfection reagents – was used as a reference. On the basis of our results, we suggest that an additional transfection barrier impairs transfection efficiency, that is: low lipoplex concentration at the cell surface. Based on the acquired knowledge we propose an optimized transfection protocol that allowed us to efficiently transfect DND41, JURKAT, MOLT3, P12-ICHIKAWA, ALL-SILL, TALL-1 human T-cell acute lymphoblastic leukemia (T-ALL) cell lines known to be difficult-to-transfect by using non-viral vectors and where LFN-based technologies fail to give satisfactory results.

Manipulation of lipoplex concentration at the cell surface boosts transfection efficiency in hard-to-transfect cells / Palchetti, Sara; Pozzi, Daniela; Marchini, Cristina; Amici, Augusto; Andreani, Cristina; Bartolacci, Caterina; Digiacomo, Luca; Gambini, Valentina; Cardarelli, Francesco; Di Rienzo, Carmine; Peruzzi, Giovanna; Amenitsch, Heinz; Palermo, Rocco; Screpanti, Isabella; Caracciolo, Giulio. - In: NANOMEDICINE. - ISSN 1549-9634. - STAMPA. - 13:2(2017), pp. 681-691. [10.1016/j.nano.2016.08.019]

Manipulation of lipoplex concentration at the cell surface boosts transfection efficiency in hard-to-transfect cells

PALCHETTI, SARA;POZZI, DANIELA;Digiacomo, Luca;CARDARELLI, FRANCESCO;Palermo, Rocco;SCREPANTI, Isabella;CARACCIOLO, Giulio
2017

Abstract

To date, efficiency upon non-viral DNA delivery remains low and this implies the existence of unidentified transfection barriers. Here we explore the mechanisms of action of multicomponent (MC) cationic liposome/DNA complexes (lipoplexes) by a combination of reporter technologies, dynamic light scattering (DLS), synchrotron small angle X-ray scattering (SAXS), fluorescence activated cell sorting (FACS) analysis and laser scanning confocal microscopy (LSCM) in live cells. Lipofectamine – the gold standard among transfection reagents – was used as a reference. On the basis of our results, we suggest that an additional transfection barrier impairs transfection efficiency, that is: low lipoplex concentration at the cell surface. Based on the acquired knowledge we propose an optimized transfection protocol that allowed us to efficiently transfect DND41, JURKAT, MOLT3, P12-ICHIKAWA, ALL-SILL, TALL-1 human T-cell acute lymphoblastic leukemia (T-ALL) cell lines known to be difficult-to-transfect by using non-viral vectors and where LFN-based technologies fail to give satisfactory results.
2017
Cationic liposomes; Cell transfection; Hard-to-transfect cells; Lipoplexes; Transfection efficiency; Medicine (miscellaneous); Bioengineering; Biomedical Engineering; Molecular Medicine; Materials Science (all); 3003
01 Pubblicazione su rivista::01a Articolo in rivista
Manipulation of lipoplex concentration at the cell surface boosts transfection efficiency in hard-to-transfect cells / Palchetti, Sara; Pozzi, Daniela; Marchini, Cristina; Amici, Augusto; Andreani, Cristina; Bartolacci, Caterina; Digiacomo, Luca; Gambini, Valentina; Cardarelli, Francesco; Di Rienzo, Carmine; Peruzzi, Giovanna; Amenitsch, Heinz; Palermo, Rocco; Screpanti, Isabella; Caracciolo, Giulio. - In: NANOMEDICINE. - ISSN 1549-9634. - STAMPA. - 13:2(2017), pp. 681-691. [10.1016/j.nano.2016.08.019]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/932442
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