Role of cholesterol on the transfection barriers of cationic lipid/DNA complexes

TY - JOUR

T1 - Role of cholesterol on the transfection barriers of cationic lipid/DNA complexes

AU - Pozzi, Daniela

AU - Cardarelli, Francesco

AU - Salomone, Fabrizio

AU - Marchini, Cristina

AU - Amenitsch, Heinz

AU - Barbera, Giorgia La

AU - Caracciolo, Giulio

PY - 2014/8/18

Y1 - 2014/8/18

N2 - 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 1,2-dioleoyl-3-trimethylammonium-propane and dioleoylphosphocholine in Chinese hamster ovary cells. The transfection mechanisms of cholesterol-containing lipoplexes have been investigated by TE, synchrotron small angle X-ray scattering, and laser scanning confocal microscopy experiments. We prove that cholesterol-containing lipoplexes enter the cells using different endocytosis pathways. Formulations with high cholesterol content efficiently escape from endosomes and exhibit a lamellar-nonlamellar phase transition in mixture with biomembrane mimicking lipid formulations. This might explain both the DNA release ability and the high transfection efficiency. These studies highlight the enrichment in cholesterol as a decisive factor for transfection and will contribute to the rational design of lipid nanocarriers with superior TE.

AB - 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 1,2-dioleoyl-3-trimethylammonium-propane and dioleoylphosphocholine in Chinese hamster ovary cells. The transfection mechanisms of cholesterol-containing lipoplexes have been investigated by TE, synchrotron small angle X-ray scattering, and laser scanning confocal microscopy experiments. We prove that cholesterol-containing lipoplexes enter the cells using different endocytosis pathways. Formulations with high cholesterol content efficiently escape from endosomes and exhibit a lamellar-nonlamellar phase transition in mixture with biomembrane mimicking lipid formulations. This might explain both the DNA release ability and the high transfection efficiency. These studies highlight the enrichment in cholesterol as a decisive factor for transfection and will contribute to the rational design of lipid nanocarriers with superior TE.

UR - http://www.scopus.com/inward/record.url?scp=84929430382&partnerID=8YFLogxK

U2 - 10.1063/1.4892915

DO - 10.1063/1.4892915

M3 - Journal article

AN - SCOPUS:84929430382

SN - 0003-6951

VL - 105

JO - Applied Physics Letters

JF - Applied Physics Letters

IS - 7

M1 - 073701

ER -