Improved lipid-mediated gene transfer into primary cultures of hippocampal neurons.
We have examined lipids as transfection agents to introduce recombinant plasmids into primary cultures of rat hippocampal neurons. By modifying the protocol for transfection mediated by the commercial reagent DOTAP, we were able to achieve a transfection efficiency of about 3%. Expression of various transfected gene products was sustained for several weeks in culture, the neurons developed normally and the transfected gene products were targeted to the appropriate subcellular compartment.
Kaech
S
S
Laboratory of Neurobiology, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892-4062, USA.
Kim
J B
JB
Cariola
M
M
Ralston
E
E
D000818:Animals; D002411:Cation Exchange Resins; D002478:Cells, Cultured; D004247:DNA / Q000008:administration & dosage* / Q000378:metabolism; D004337:Drug Carriers; D005229:Fatty Acids, Monounsaturated; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D005998:Glycine / Q000031:analogs & derivatives; D006624:Hippocampus / Q000166:cytology*; D007145:Immunoglobulin kappa-Chains / Q000032:analysis / Q000096:biosynthesis; D008055:Lipids; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D010957:Plasmids; D000644:Quaternary Ammonium Compounds; D051381:Rats; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis*; D013096:Spermine / Q000031:analogs & derivatives; D014162:Transfection / Q000379:methods*
D002411:Cation Exchange Resins; D004337:Drug Carriers; D005229:Fatty Acids, Monounsaturated; D005456:Fluorescent Dyes; D007145:Immunoglobulin kappa-Chains; D008055:Lipids; C086724:Lipofectamine; D000644:Quaternary Ammonium Compounds; D011994:Recombinant Proteins; C061134:dioctadecylamidoglycylspermine; D013096:Spermine; D004247:DNA; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane; D005998:Glycine