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Comparing the caveolae mediated endocytosis of two DNA-chitosan polyplexes
Norwegian University of Science and Technology, Faculty of Natural Sciences and Technology, Department of Physics.
2011 (English)MasteroppgaveStudent thesis
Abstract [en]

Understanding intracellular processing of gene vectors will help to improve vector design in gene therapy. Chitosan nanoparticles have been previously identied as safe and non toxic gene vector. Linear chitosan oligomer (LCO) and self branched trisaccharide chitosan oligomer (SBTCO) have been shown to be able to pack DNA, balancing between complexation and intracellular unpacking . However, the transfection ecacies of these two chitosans diers considerably with the level of transgene expression higher for SBTCO compared to LCO. SBTCO have been recently reported to be taken up solely by caveolae mediated endocytosis (CvME) while LCO uses both clathrin mediated endocytosis (CME) and CvME. In the present study, the CvME was studied. An immunostaining protocol for detection of caveolin (cav) was established. Polyplexes of SBTCO seemed to trigger the formation of more caveosomes than did LCO polyplexes. SBTCO polyplexes were more localised in the caveosome than LCO polyplexes at 3 hours incubation period. These ndings suggested that SBTCO polyplexes delivers more DNA into the cell than LCO polyplexes and that SBTCO polyplexes are processed intracellularly solely via the CvME pathway. Likewise, it suggested LCO polyplexes have preferred intracellular processing pathway which is not CvME. Collectively, these results demonstrated that SBTCO is protected from the degradative endosome and might therefore be an ecient and good tool to deliver therapeutic DNA.

Place, publisher, year, edition, pages
Institutt for fysikk , 2011. , 82 p.
Keyword [no]
ntnudaim:6486, MSMEDTEK Medical Technology, Biotechnology
URN: urn:nbn:no:ntnu:diva-14200Local ID: ntnudaim:6486OAI: diva2:448547
Available from: 2011-10-17 Created: 2011-10-17

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