Lipopolyamine-mediated transfection allows gene expression studies in primary neuronal cells.
A simple and efficient transfection technique based on lipopolyamine-coated DNA that can be used for gene transfer in cerebellar granular neurons is described. Gene transfer is achieved by exposure of cells to a DNA/lipid complex obtained by simple mixing of lipopolyamine and plasmid DNA. This procedure may represent a general tool of physiological investigations in primary cells. We show that the promoters of the introduced chimera genes are regulated by their respective trans-acting factors and may be modulated via membrane receptors and second messengers. This procedure has no noticeable toxic effects, nor does it seem to interfere with complex physiological behavior like neuronal differentiation.
Loeffler
J P
JP
Institut de Physiologie, U. 309 du CNRS, Strasbourg, France.
Barthel
F
F
Feltz
P
P
Behr
J P
JP
Sassone-Corsi
P
P
Feltz
A
A
D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002531:Cerebellum / Q000166:cytology; D005786:Gene Expression Regulation; D005818:Genetic Engineering; D005998:Glycine / Q000031:analogs & derivatives* / Q000494:pharmacology; D008969:Molecular Sequence Data; D009474:Neurons / Q000502:physiology*; D010714:Phosphatidylethanolamines / Q000494:pharmacology*; D010957:Plasmids / Q000502:physiology; D015290:Second Messenger Systems; D013096:Spermine / Q000031:analogs & derivatives* / Q000494:pharmacology; D014162:Transfection
D010714:Phosphatidylethanolamines; C061134:dioctadecylamidoglycylspermine; C061135:1,2-dipalmitoylphosphatidylethanolamidospermine; C063681:dioctadecylamidospermine; D013096:Spermine; D005998:Glycine