title: Cellular uptake of DNA-chitosan nanoparticles: the role of clathrin- and caveolae-mediated pathways.
abstract: 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.
authors.affiliation: Department of Physics, The Norwegian University of Science and Technology, 7491 Trondheim, Norway.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Garaiova
authors.forename: Sabina P
authors.initials: SP
authors.lastname: Strand
authors.forename: Nina K
authors.initials: NK
authors.lastname: Reitan
authors.forename: Sylvie
authors.initials: S
authors.lastname: Lélu
authors.forename: Sigmund Ø
authors.initials: SØ
authors.lastname: Størset
authors.forename: Kristian
authors.initials: K
authors.lastname: Berg
authors.forename: Jostein
authors.initials: J
authors.lastname: Malmo
authors.forename: Oladayo
authors.initials: O
authors.lastname: Folasire
authors.forename: Astrid
authors.initials: A
authors.lastname: Bjørkøy
authors.forename: Catharina de L
authors.initials: Cde L
authors.lastname: Davies
mesh_terms: D021941:Caveolae / Q000187:drug effects / Q000378:metabolism*; D048271:Chitosan / Q000737:chemistry* / Q000378:metabolism*; D002746:Chlorpromazine / Q000494:pharmacology; D002966:Clathrin / Q000378:metabolism*; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004337:Drug Carriers / Q000737:chemistry / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects; D019833:Genistein / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D006835:Hydrazones / Q000494:pharmacology; D053758:Nanoparticles; D013499:Surface Properties; D013696:Temperature; D014162:Transfection
chemical_list: D002966:Clathrin; D004337:Drug Carriers; D006835:Hydrazones; C511794:N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide; D004247:DNA; D048271:Chitosan; D019833:Genistein; D002746:Chlorpromazine