Specific gene transfer mediated by lactosylated poly-L-lysine into hepatoma cells.
Plasmid DNA/glycosylated polylysine complexes were used to transfer in vitro a luciferase reporter gene into human hepatoma cells by a receptor-mediated endocytosis process. HepG2 cells which express a galactose specific membrane lectin were efficiently and selectively transfected with pSV2Luc/lactosylated polylysine complexes in a sugar dependent manner: i) HepG2 cells which do not express membrane lectin specific for mannose were quite poorly transfected with pSV2Luc/mannosylated polylysine complexes, ii) HeLa cells which do not express membrane lectin specific for galactose were not transfected with pSV2Luc/lactosylated polylysine complexes. The transfection efficiency of HepG2 cells with pSV2Luc/lactosylated polylysine complexes was greatly enhanced either in the presence of chloroquine or in the presence of a fusogenic peptide. A 22-residue peptide derived from the influenza virus hemagglutinin HA2 N-terminal polypeptide that mimics the fusogenic activity of the virus, was selected. In the presence of the fusogenic peptide, the luciferase activity in HepG2 cells was 10 fold larger than that of cells transfected with pSV2Luc/lactosylated polylysine complexes in the presence of chloroquine.
Département de Biochimie des Glycoconjugués et Lectines Endogènes, CNRS, Université d'Orléans, France.
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P
Midoux
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C
Mendes
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A
Legrand
J
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Raimond
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Mayer
M
M
Monsigny
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AC
Roche
D000595:Amino Acid Sequence; D006528:Carcinoma, Hepatocellular; D002738:Chloroquine; D019793:Fluorescein; D005452:Fluoresceins; D006031:Glycosylation; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D006801:Humans; D007785:Lactose; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D010957:Plasmids / Q000378:metabolism; D011107:Polylysine; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured; D014760:Viral Fusion Proteins
D005452:Fluoresceins; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D014760:Viral Fusion Proteins; D011107:Polylysine; D002738:Chloroquine; D007785:Lactose; D019793:Fluorescein