Optimization of liposome mediated transfection of a neuronal cell line.
A cell line derived from sensory neurons was transfected with high efficiency using cationic liposomes, formulated from 3 beta [N-(N',N'-dimethylaminoethane)carbamoyl]-cholesterol (DC-Chol) and dioleoyl L-alpha-phosphatidylethanolamine (DOPE). This is the first time that cationic liposomes of this type have been reported to transfect a neuronal cell line. We used a reporter gene construct expressing beta-galactosidase under the control of the cytomegalovirus immediate early promoter and routinely observed transfection efficiencies > 40%. Parameters affecting transfection efficiency were examined and the ratio of DNA to liposome proved to be crucial. Liposome formulation procedures and cell transfection protocols devised here will be used as a basis for further in vivo and in vitro work.
McQuillin
A
A
Department of Psychiatry and Behavioural Sciences, UCL Medical School, Windeyer Institute of Medical Sciences, London, UK.
Murray
K D
KD
Etheridge
C J
CJ
Stewart
L
L
Cooper
R G
RG
Brett
P M
PM
Miller
A D
AD
Gurling
H M
HM
D000818:Animals; D002412:Cations; D002460:Cell Line; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000737:chemistry; D017930:Genes, Reporter; D008081:Liposomes; D009475:Neurons, Afferent / Q000502:physiology*; D010714:Phosphatidylethanolamines / Q000737:chemistry*; D051381:Rats; D014162:Transfection / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D002412:Cations; D008081:Liposomes; D010714:Phosphatidylethanolamines; C070648:3-(N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol; C094877:dioleoyl phosphatidylethanolamine; D002784:Cholesterol; D001616:beta-Galactosidase