Influence of membrane-active peptides on lipospermine/DNA complex mediated gene transfer.
To explore whether endosomal release presents a major barrier to lipospermine-mediated gene delivery, acidic membrane-active peptides derived from influenza virus or artificial sequences were incorporated into DNA/dioctadecylamidoglycylspermine (= Transfectam) complexes. Depending on the cell line used, gene expression levels are approximately 3-30-fold higher than those obtained by applying DNA complexed to optimal amounts of Transfectam alone. In addition, gene transfer efficiency of DNA complexes with lower amounts of Transfectam (1.5-2 charge equiv) is increased by a factor of up to 1000 by peptides INF6 (influenza virus derived sequence) and INF10 (artificial sequence). The helper lipids 1,2-dioleoylphosphatidylethanolamine, egg phosphatidylethanolamine, and 1,2-dioleoyl-racglycerol also can enhance the gene transfer. Thus, endosomal escape seems to be only a moderate barrier for optimized, positively charged DNA/Transfectam complexes, but a substantial bottleneck for less positively charged complexes.
Kichler
A
A
Institute of Biochemistry, University of Vienna, Austria.
Mechtler
K
K
Behr
J P
JP
Wagner
E
E
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics* / Q000378:metabolism; D004337:Drug Carriers; D004705:Endocytosis; D018014:Gene Transfer Techniques; D005998:Glycine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000378:metabolism; D006801:Humans; D008081:Liposomes; D008565:Membrane Proteins / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000008:administration & dosage / Q000378:metabolism; D013096:Spermine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000378:metabolism
D004337:Drug Carriers; D008081:Liposomes; D008565:Membrane Proteins; D010455:Peptides; D010714:Phosphatidylethanolamines; C061134:dioctadecylamidoglycylspermine; D013096:Spermine; C020888:1,2-dielaidoylphosphatidylethanolamine; D004247:DNA; D005998:Glycine