Opposing roles of syndecan-1 and syndecan-2 in polyethylenimine-mediated gene delivery

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DOIResolve DOI: http://doi.org/10.1074/jbc.M705424200
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TypeArticle
Journal titleJournal of Biological Chemistry
Volume283
Issue12
Pages76977704; # of pages: 18
SubjectAnalysis; animal; bio; biotechnology; cells; endocytosis; gene expression; incomplet; proteoglycans; tail; transfection; polyethyleneimine; clustering
AbstractPolyethylenimines (PEIs) are efficient non-viral vectors for gene transfer. Heparan sulphate proteoglycans have been proposed to be the cell surface receptors for PEI-DNA complexes (polyplexes). Here, we investigated if syndecan-1 (SDC1) and syndecan-2 (SDC2) may be involved in PEI-mediated transfection. Following addition of polyplexes to HEK293 cells, GFP-tagged SDCs rapidly formed clusters with PEI that were dependent of lipid rafts integrity. However, while SDC1 overexpression slightly enhanced PEImediated gene expression, SDC2 dramatically inhibited it. Confocal microscopy analysis showed that SDC1:polyplex endocytosis occurs within minutes after addition of polyplexes while that of SDC2:polyplexes takes hours. Expression of SDC1 cytoplasmic deletion mutants revealed that the SDC1 cytoplasmic tail is required for gene expression, but not for clustering or endocytosis, while overexpression of SDC1/SDC2 chimeras showed that SDC2 ectodomain is responsible for the inhibitory effect on gene transfer. Taken together, this study provides evidence that SDCs may have opposing effects on PEI-mediated transfection.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada; NRC Biotechnology Research Institute
NoteDA - 20080124IS - 0021-9258 (Print)LA - ENGPT - JOURNAL ARTICLE
Peer reviewedNo
NRC number47810
47810
NPARC number8925865
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Record identifierd3e4fdc5-a460-4644-b40c-5de321a324a3
Record created2009-04-23
Record modified2016-05-09
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