Expression of hepatitis B virus surface antigen in adult rat liver. Co-introduction of DNA and nuclear protein by a simplified liposome method.
We established a simple and efficient method for gene transfer in vitro (to cultured cells) and in vivo (to an adult organ) using liposomes. Plasmid DNA and proteins were efficiently co-encapsulated in liposomes by agitation and sonication, and were co-introduced into cells by hemagglutinating virus of Japan (HVJ)-mediated membrane fusion. Introduction of the Escherichia coli beta-galactosidase gene with non-histone chromosomal protein high mobility group 1 (HMG1) into LLCMK2 cells resulted in about 3 times higher beta-galactosidase activity than that on introduction of the gene alone. Two days after injection of HVJ-liposomes containing the beta-galactosidase gene and HMG1 under the perisplanchnic membrane of adult rat liver, hepatic cells near the injection site were found by 5-bromo-4-chloro-3-indolyl beta-D-galactoside staining to have beta-galactosidase activity. After similar injection of HVJ-liposomes containing the hepatitis B virus surface antigen (HBsAg) gene and HMG1, HBsAg was detected in the serum for 9 days with a maximum of 25-45 ng/ml on day 2 after the injection.
Kato
K
K
Institute for Molecular and Cellular Biology, Osaka University, Japan.
Nakanishi
M
M
Kaneda
Y
Y
Uchida
T
T
Okada
Y
Y
D000818:Animals; D004247:DNA; D005814:Genes, Viral; D006514:Hepatitis B Surface Antigens / Q000032:analysis* / Q000097:blood; D006609:High Mobility Group Proteins / Q000235:genetics; D007124:Immunoenzyme Techniques; D008081:Liposomes; D008099:Liver / Q000201:enzymology / Q000276:immunology / Q000382:microbiology*; D010222:Parainfluenza Virus 1, Human / Q000235:genetics; D010957:Plasmids; D051381:Rats; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
D006514:Hepatitis B Surface Antigens; D006609:High Mobility Group Proteins; D008081:Liposomes; D004247:DNA; D001616:beta-Galactosidase