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.
|Titolo:||Manipulation of lipoplex concentration at the cell surface boosts transfection efficiency in hard-to-transfect cells|
|Data di pubblicazione:||2017|
|Parole Chiave:||Cationic liposomes; Cell transfection; Hard-to-transfect cells; Lipoplexes; Transfection efficiency; Animals; Cell Line; DNA; Humans; Lipids; Scattering, Small Angle; X-Ray Diffraction; Liposomes; Transfection|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1016/j.nano.2016.08.019|
|Appare nelle tipologie:||1.1 Articolo in rivista|