Intracellular Trafficking and Decondensation Kinetics of Chitosan–pDNA Polyplexes

摘要: The transfection efficiency (TE) of chitosan–plasmid DNA (pDNA) polyplexes can be critically modulated by the polymer degree deacetylation (DDA) and molecular weight (MW). This study was performed to test hypothesis that TE dependence on chitosan MW DDA is related polyplex stability, hence their intracellular decondensation/unpacking kinetics. Major barriers nonviral gene transfer were studied image-based quantification. Although uptake increased with DDA, it did not appear a structure-dependent process affecting TE, nor nuclear entry. Colocalization analysis showed all chitosans trafficked through lysosomes similar Fluorescent resonant energy (FRET) revealed distinct relationship between dissociation rate. most efficient an intermediate stability kinetics dissociation, which occurred in synchrony lysosomal escape. In contrast, rapid before escape found for inefficient low whereas highly stable complex formed high dissociate even after 24 hours. identified decondensation relation critical polyplexes.