Enhanced transgene expression in rat brain cell cultures with a disulfide-containing cationic lipid

The transfection efficiency of a disulfide-containing cationic lipid, 1',2' dioleoyl-sn-glycero-3'-succinyl-2-hydroxyethyl disulfide ornithine conjugate (DOGSDSO) and its non-disulfide analog (DOGSHDO) were compared in neuronal, astroglial and microglial cultures from newborn rat cerebral cortex. We hypothesized that the relatively high intracellular concentrations of reductive substances in the cytoplasm may help to cleave the reversible disulfide bond in DOGSDSO, thus increasing free DNA and decreasing toxicity due to rapid degradation of the lipid. We have demonstrated through mass spectrometric analysis that a reductive compound, e.g. dithiothreitol (DTT) could degrade the disulfide lipid. DOGSDSO was more efficient at transfecting each type of brain cell than were the non-disulfide DOGSHDO and DOTAP (1,2-dioleoyl-3-trimethyl-ammonium-propane) liposomes. These results demonstrate that disulfide-containing cationic liposomes facilitate gene transfection in cultured rat brain cells.