Cellular uptake of DNA–chitosan nanoparticles: The role of clathrin- and caveolae-mediated pathways
2012
Garaiova, Zuzana | Strand, Sabina P. | Reitan, Nina K. | Lélu, Sylvie | Størset, Sigmund Ø | Berg, Kristian | Malmo, Jostein | Folasire, Oladayo | Bjørkøy, Astrid | de L. Davies, Catharina
The success of gene therapy depends on efficient delivery of DNA and requires a vector. A promising non-viral vector is chitosan. We tailored chitosan to optimize it for transfection by synthesizing self-branched and trisaccharide-substituted chitosan oligomers (SBTCO), which show superior transfection efficacy compared with linear chitosan (LCO). The aim of the work was to compare the cellular uptake and endocytic pathways of polyplexes formed by LCO and SBTCO. Both polyplexes were taken up by the majority of the cells, but the uptake of LCO was lower than SBTCO polyplexes. LCO polyplexes were internalized through both clathrin-dependent and clathrin-independent pathways, whereas SBTCO polyplexes were primarily taken up by clathrin-independent endocytosis. The different level of cellular uptake and the distinct endocytic pathways, may explain the difference in transfection efficacy. This was supported by the observation that photochemical internalization increased the transfection by LCO polyplexes considerably, whereas no effect on transfection was found for SBTCO polyplexes.
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