Cytoskeletal involvement in the cellular trafficking of plasmid/PEI derivative complexes

Development of a recombinant fusion protein based on the dynein light chain LC8 for non-viral gene delivery

“…The results indicate that pDNA:LD4 complexes strongly rely on the cells cytoskeleton for intracellular trafficking and pDNA delivery. However, the decrease in transfection efficiency caused by the disruption of the cytoskeleton has already been described in the literature using pDNA:Lipofectamine™ and pDNA:PEI (polyethyleneimine) complexes [39,40]. Despite microtubules tend to facilitate intracellular trafficking of vectors via active retrograde transport of the vesicles formed after endocytosis, they can also contribute to their degradation since most of the vectors remain entrapped inside late endosomes and lysosomes and are finally destroyed in these vesicles [41,42].…”

“…Despite microtubules tend to facilitate intracellular trafficking of vectors via active retrograde transport of the vesicles formed after endocytosis, they can also contribute to their degradation since most of the vectors remain entrapped inside late endosomes and lysosomes and are finally destroyed in these vesicles [41,42]. On the other hand, the observed decrease in transfection efficiency when cells were pre-treated with cytochalasin D can be credited to the role attributed to actin filaments in the early steps of complex entry in the cell [40] as they are important to receptormediated endocytosis and might be involved in other pathways of internalization [43]. Disruption of actin filaments tends to decrease the internalization of complexes and hence lower transfection efficiency.…”

Junk DNA enhances pEI-based non-viral gene delivery

“…Among the best characterized and most frequently used polymeric carriers are the polyethylenimines (pEI), which are generally considered as a golden standard for nonviral gene transfection. pEI-based transfection was first described by Boussif et al (1995) and since then numerous efforts have been made to gain insights into the mechanism of action (Akinc and Langer, 2002;Boeckle et al, 2004;Breunig et al, 2006;Breuzard et al, 2008;Brunner et al, 2002;Clamme et al, 2003;Erbacher et al, 2004;Forrest and Pack, 2002;Grosse et al, 2007;Itaka et al, 2004;Ogris et al, 1998;Rejman et al, 2006;Godbey et al, 1999b,d) and to further optimize this polymer (Godbey et al, 1999a,c;Neu et al, 2005;Ogris et al, 1999;Dunlap et al, 1997;Kircheis et al, 2001).…”

Synergistic effects between natural histone mixtures and polyethylenimine in non-viral gene delivery in vitro

“…Fluorescein-labelled pDNA (1.4 g) was again complexed by histones and PEI adjusted to the corresponding pDNA concentration and incubated with cells for 2 days. Cells were rinsed twice with sodium citrate buffer (25.5 mM citric acid, 24.5 mM sodium citrate, 280 mM sucrose, 0.01 mM deferoxamine mesylate, pH 4.6) to remove cell surface-bound particles (Ghosh et al, 1994;Grosse et al, 2007), followed by two washing steps with PBS. Cells were fixed with 0.5 ml 3.7% formaldehyde, washed with PBS and subsequently stained with 4 -6-diamidino-2-phenylindole (DAPI) for 15 min.…”