Interaction of oligonucleotides with cationic lipids: the relationship between electrostatics, hydration and state of aggregation.
Lipoplexes, which are spontaneously formed complexes between oligonucleotide (ODN) and cationic lipid, can be used to deliver ODNs into cells, both in vitro and in vivo. The present study was aimed at characterizing the interactions associated with the formation of lipoplexes, specifically in terms of electrostatics, hydration and particle size. Large unilamellar vesicles (approximately 100 nm diameter), composed of either DOTAP, DOTAP/cholesterol (mole ratio 1:1) or DOTAP/DOPE (mole ratio 1:1) were employed as a model of cationic liposomes. Neutral vesicles ( approximately 100 nm diameter), composed of DOPC/DOPE (mole ratio 1:1), were employed as control liposomes. After ODN addition to vesicles, at different mole ratios, changes in pH and electrical surface potential at the lipid-water interface were analyzed by using the fluorophore heptadecyl-7-hydroxycoumarin. In separate 'mirror image' experiments, liposomes were added at different mole ratios to fluorescein isothiocyanate-labeled ODNs, thus yielding data about changes in the pH near the ODN molecules induced by the complexation with the cationic lipid. Particle size distribution and turbidity fluctuations were analyzed by the use of photon correlation spectroscopy and static light-scattering, respectively. In additional fluorescent probe studies, TMADPH was used to quantify membrane defects while laurdan was used to measure the level of hydration at the water-lipid interface. The results indicate that mutual neutralization of cationic lipids by ODNs and vice versa is a spontaneous reaction and that this neutralization is the main driving force for lipoplex generation. When lipid neutralization is partial, induced membrane defects cause the lipoplexes to exhibit increased size instability.
Meidan
V M
VM
Department of Biochemistry, Hadassah Medical School, The Hebrew University, P.O. Box 12272, Jerusalem, Israel.
Cohen
J S
JS
Amariglio
N
N
Hirsch-Lerner
D
D
Barenholz
Y
Y
D015081:2-Naphthylamine / Q000031:analogs & derivatives; D002412:Cations; D002784:Cholesterol / Q000737:chemistry; D004161:Diphenylhexatriene / Q000031:analogs & derivatives; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D007848:Laurates; D008055:Lipids / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry; D020319:Oligodeoxyribonucleotides, Antisense / Q000737:chemistry; D009841:Oligonucleotides / Q000737:chemistry*; D010316:Particle Size; D010714:Phosphatidylethanolamines / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D055672:Static Electricity; D013873:Thionucleotides / Q000737:chemistry; D014468:Umbelliferones
D002412:Cations; D005229:Fatty Acids, Monounsaturated; D005456:Fluorescent Dyes; D007848:Laurates; D008055:Lipids; D008081:Liposomes; D020319:Oligodeoxyribonucleotides, Antisense; D009841:Oligonucleotides; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D013873:Thionucleotides; D014468:Umbelliferones; C095188:hybrid oligonucleotide phosphorothioate; D004161:Diphenylhexatriene; C031827:4-heptadecylumbelliferone; C033628:1-(4-(trimethylamino)phenyl)-6-phenylhexa-1,3,5-triene; C020888:1,2-dielaidoylphosphatidylethanolamine; D002784:Cholesterol; D015081:2-Naphthylamine; D016650:Fluorescein-5-isothiocyanate; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane; C065580:laurdan