Neuronal gene delivery by negatively charged pullulan-spermine/DNA anioplexes.

摘要: Nonviral gene transfer to neurons remains unreliable due a lack of effective and nontoxic vectors. Here, we achieved neuronal delivery through salt-free complexation plasmid DNA pullulan-spermine, conjugate prepared from naturally derived polysaccharide polyamine. Specifically, at low spermine nitrogen:DNA phosphate (N:P) ratios, complexes formed with zeta-potential diameter approximately-40mV 350nm, respectively. Higher N:P ratios increased the approximately +10mV. All were stable for least 1 week protected degradation. In vitro transfection rat sensory occurred all but uniquely, efficiency was highest anionic (anioplexes). Subsequent analyses revealed inhibition reporter expression by asialofetuin (1mg/ml) methyl-beta-cyclodextrin (5mm), indicating utilization glycoprotein-specific interactions lipid rafts uptake intracellular trafficking. marked contrast commercial cationic reagent, anioplexes did not exhibit measurable cytotoxicity up 20microg/ml DNA. Additionally, maintained in presence serum antibiotics. Based on these favorable properties, successfully established two methods cultured adult tissue explants. Collectively, data suggest that negatively charged pullulan-spermine/DNA could represent an technology, particularly neurons.