Most lipid formulations need cholesterol for efficient transfection, but the precise motivation remains unclear. Here, we have investigated the effect of cholesterol on the transfection efficiency (TE) of cationic liposomes made of l, 2-dioleoyl-3-trimethylammonium-propane (DOTAP) and dioleoylphosphocholine (DOPC) in Chinese hamster ovary cells. The transfection mechanisms of cholesterol-containing lipoplexes have been investigated by TE, synchrotron small angle X-ray scattering (SAXS) and laser scanning confocal microscopy (LSCM) experiments. We prove that cholesterol-containing lipoplexes enter the cells using different endocytosis pathways. Formulations with high cholesterol content efficienlty escape from endosomes. These studies will contribute to the rational design of novel lipid nanocarriers with superior TE.
Effect of cholesterol on the transfection efficiency of DOTAP-containing lipoplexes
Cardarelli, Francesco;Salomone, Fabrizio;
2013
Abstract
Most lipid formulations need cholesterol for efficient transfection, but the precise motivation remains unclear. Here, we have investigated the effect of cholesterol on the transfection efficiency (TE) of cationic liposomes made of l, 2-dioleoyl-3-trimethylammonium-propane (DOTAP) and dioleoylphosphocholine (DOPC) in Chinese hamster ovary cells. The transfection mechanisms of cholesterol-containing lipoplexes have been investigated by TE, synchrotron small angle X-ray scattering (SAXS) and laser scanning confocal microscopy (LSCM) experiments. We prove that cholesterol-containing lipoplexes enter the cells using different endocytosis pathways. Formulations with high cholesterol content efficienlty escape from endosomes. These studies will contribute to the rational design of novel lipid nanocarriers with superior TE.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
