The role of cholesterol and structurally related molecules in enhancing transfection of cationic liposome-DNA complexes.
Motivated by its important role in gene delivery, we have studied the effect of cholesterol and analogs on the transfection efficiency (TE) of lamellar cationic liposome-DNA (CL-DNA) complexes in vitro. Addition of cholesterol to low-transfecting DOTAP/DOPC-DNA complexes increases TE, with 15 mol % cholesterol already yielding 10-fold improvement. Steroids lacking the alkyl tail only modestly enhance TE, while molecules retaining it strongly enhance TE. All steroid-containing CL-DNA complexes exhibit the lamellar structure. The increase in experimentally determined membrane charge density (a universal parameter governing the TE of lamellar CL-DNA complexes) with cholesterol content alone cannot account for the rapid increase of TE. Instead, the reduction of the hydration repulsion layer of the membrane, caused by replacement of DOPC by cholesterol, promotes fusion between cationic membranes of CL-DNA complexes and anionic endosomal membranes, thus facilitating release of complexes and enhancing TE.
Zidovska
Alexandra
A
Materials, Physics, and Molecular, Cellular and Developmental Biology Departments, University of California at Santa Barbara, Santa Barbara, California 93106, USA.
Evans
Heather M
HM
Ahmad
Ayesha
A
Ewert
Kai K
KK
Safinya
Cyrus R
CR
D002412:Cations / Q000737:chemistry; D002784:Cholesterol / Q000737:chemistry* / Q000494:pharmacology; D004247:DNA / Q000737:chemistry*; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry*; D015394:Molecular Structure; D010713:Phosphatidylcholines / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D053838:Scattering, Small Angle; D014162:Transfection / Q000379:methods*; D014961:X-Ray Diffraction
D002412:Cations; D005229:Fatty Acids, Monounsaturated; D010713:Phosphatidylcholines; D000644:Quaternary Ammonium Compounds; D004247:DNA; D002784:Cholesterol; C017251:1,2-oleoylphosphatidylcholine; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane