Cationic liposome-mediated intravenous gene delivery.
Systemic gene transfer provides new opportunities for the analysis of gene function and gene regulation in vivo, as well as for human gene therapy. We used the chloramphenicol acetyltransferase reporter gene to examine several parameters important for the development of efficient, cationic liposome-mediated, intravenous (IV) gene transfer in mice. We then demonstrated that this approach can produce high level expression of biologically important genes. Specifically, we assessed the relationship of expression vector design to the level of systemic gene expression produced, and compared transfection levels produced by intravenously injecting DNA alone versus DNA-liposome complexes. We found that both the position of the heterologous intron, and the promoter element used in the expression plasmid, significantly affected the level of systemic gene expression produced. Although intravenous injection of plasmid DNA alone transfected every tissue analyzed, liposome-mediated delivery was much more efficient. We also established that repeated i.v. injection of DNA-liposome complexes produced high level systemic transfection. The second injection of DNA-liposome complexes produced levels of gene expression at least as high as those following a single i.v. injection. Thus, unlike some viral vectors, a neutralizing host-immune response does not limit re-expression, following reinjection of DNA-liposome complexes. Finally, we showed that the expression vectors which produced the highest levels of chloramphenicol acetyltransferase reporter gene expression could also produce high level expression of two colony stimulating factor genes in mice. Specifically, i.v. injection of liposomes complexed to expression vectors into which we had inserted either the murine granulocyte-macrophage-colony stimulating factor cDNA or the human granulocyte-CSF cDNA, produced circulating levels of the corresponding colony stimulating factor gene product comparable to levels which have been shown previously to be both biologically and therapeutically significant.
Liu
Y
Y
California Pacific Medical Research Institute, San Francisco, California 94115, USA.
Liggitt
D
D
Zhong
W
W
Tu
G
G
Gaensler
K
K
Debs
R
R
D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000008:administration & dosage*; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D016179:Granulocyte Colony-Stimulating Factor / Q000097:blood / Q000235:genetics; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000097:blood / Q000235:genetics; D006801:Humans; D007275:Injections, Intravenous; D008081:Liposomes / Q000008:administration & dosage*; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic
D008081:Liposomes; D016179:Granulocyte Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D004247:DNA