Efficient gene transfer into mammalian cells using fusogenic liposome.
Fusogenic liposome (FL) based on Sendai virus constitutes a unique system that delivers the content efficiently into animal cells in vitro and in vivo. In this study we characterized unilamellar FL as a gene transfer vector in comparison with cationic lipid (CL)-DNA complex. FL transferred genes efficiently into cultured cells even when incubated for as little as 10 min, while CL-DNA complex required at least 30 min to reach the same level of gene expression. FL was also much less cytotoxic than CL-DNA complex under the conditions that resulted in the same level of gene expression. In addition, FL maintained 70% of the transfection activity even in the presence of 40% fetal calf serum (FCS), while CL-DNA complex almost completely lost their activity in the presence of 5% FCS. Furthermore, we found that FL could introduce and express luciferase gene into mouse ascites tumor cells in vivo, but CL-DNA complex could not even at higher concentrations of DNA. We conclude that unilamellar FL is a unique and efficient nonviral vector for gene transfer in vitro and in vivo.
Mizuguchi
H
H
Faculty of Pharmaceutical Science, Osaka University, Japan.
Nakagawa
T
T
Nakanishi
M
M
Imazu
S
S
Nakagawa
S
S
Mayumi
T
T
D000199:Actins / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D002470:Cell Survival; D002478:Cells, Cultured; D002645:Chickens; D018014:Gene Transfer Techniques; D005814:Genes, Viral; D008081:Liposomes; D008156:Luciferases / Q000032:analysis / Q000096:biosynthesis; D008297:Male; D008322:Mammals; D008561:Membrane Fusion; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D010222:Parainfluenza Virus 1, Human; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis*; D015183:Restriction Mapping; D012513:Sarcoma, Experimental / Q000473:pathology; D014162:Transfection / Q000379:methods*
D000199:Actins; D008081:Liposomes; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D008156:Luciferases