title: Delivery of plasmid DNA into mammalian cell lines using pH-sensitive liposomes: comparison with cationic liposomes.
abstract: We compare the transfection efficiency of plasmid DNA encoding either luciferase or beta-galactosidase encapsulated in pH-sensitive liposomes or non-pH-sensitive liposomes or DNA complexed with cationic liposomes composed of dioleoyloxypropyl-trimethylammonium:dioleoylphosphatidyl-eth anolamine (1:1, w/w) (Lipofectin) and delivered into various mammalian cell lines. Cationic liposomes mediate the highest transient transfection level in all cell-lines examined. pH-sensitive liposomes, composed of cholestryl hemisuccinate and dioleoylphosphatidylethanolamine at a 2:1 molar ratio, mediate gene transfer with efficiencies that are 1 to 30% of that obtained with cationic liposomes, while non-pH-sensitive liposome compositions do not induce any detectable transfection. Cationic liposomes mediate a more rapid uptake of plasmid DNA, to about an eightfold greater level than that obtained with pH-sensitive liposomes. The higher uptake of DNA mediated by Lipofectin accounts for part of its high transfection efficiency. Treatment of cells with chloroquine, ammonium chloride, or monensin decreases (threefold) transfection using pH-sensitive liposomes and either has no effect on or enhances cationic liposome-mediated transfection. Therefore plasma membrane fusion is not the only mechanism available to cationic liposomes; in certain cell lines DNA delivery via endocytosis is a possible parallel pathway and could augment the superior transfection efficiency observed with cationic liposomes.
authors.affiliation: School of Pharmacy, University of California, San Francisco 94143-0446.
authors.forename: J Y
authors.initials: JY
authors.lastname: Legendre
authors.forename: F C
authors.initials: FC
authors.lastname: Szoka
mesh_terms: D002460:Cell Line; D004247:DNA / Q000235:genetics*; D005786:Gene Expression Regulation; D006651:Histocytochemistry; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D007700:Kinetics; D008081:Liposomes / Q000737:chemistry*; D008156:Luciferases / Q000235:genetics; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D007202:Indicators and Reagents; D008081:Liposomes; D010714:Phosphatidylethanolamines; C020888:1,2-dielaidoylphosphatidylethanolamine; D004247:DNA; D008156:Luciferases; D001616:beta-Galactosidase