Cationic multilamellar liposome-mediated gene transfer into primary myoblasts.
To transfer foreign genes into myoblasts in primary culture, we found cationic multilamellar liposomes to be a useful mediator. When the cells were transfected with 2 micrograms of the plasmid pRSV-luc encapsulated into 50 nmol of our cationic multilamellar liposomes that had been prepared from N-(alpha-trimethylammonioacetyl)-didodecyl-D-glutamate chloride, dilauroyl phosphatidylcholine, and dioleoyl phosphatidylethanolamine in a molar ratio of 1:2:2, luciferase was expressed with high efficiency without cytotoxicity. When the cells were transfected with the plasmid pRSV-lacZ encapsulated into the same liposomes, 0.7% of the cultured myoblasts expressed beta-galactosidase without cytotoxicity. This is the first successful instance of introducing foreign genes into primary cultures of myoblasts by means of liposomes.
Institute of Applied Biochemistry, Yagi Memorial Park, Gifu, Japan.
H
H
Kojima
N
N
Ohishi
M
M
Takamori
K
K
Yagi
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D004337:Drug Carriers; D018014:Gene Transfer Techniques; D005971:Glutamates; D008081:Liposomes; D008156:Luciferases / Q000032:analysis / Q000096:biosynthesis; D018482:Muscle, Skeletal / Q000378:metabolism*; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010957:Plasmids; D051381:Rats; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D014162:Transfection / Q000379:methods*
D004337:Drug Carriers; D005971:Glutamates; D008081:Liposomes; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D011994:Recombinant Proteins; C084512:didodecyl N-(alpha-trimethylammonioacetyl)glutamate; C015153:1,2-dilauroylphosphatidylcholine; C020888:1,2-dielaidoylphosphatidylethanolamine; D008156:Luciferases