TY - JOUR
T1 - Structural analysis of DNA complexation with cationic lipids
AU - Marty, Regis
AU - N'soukpoé-Kossi, Christophe N.
AU - Charbonneau, David
AU - Weinert, Carl Maximilian
AU - Kreplak, Laurent
AU - Tajmir-Riahi, Heidar Ali
N1 - Funding Information:
Natural Sciences and Engineering Research Council of Canada (NSERC). Funding for open access charge has been waived by Oxford University Press.
PY - 2009
Y1 - 2009
N2 - Complexes of cationic liposomes with DNA are promising tools to deliver genetic information into cells for gene therapy and vaccines. Electrostatic interaction is thought to be the major force in lipid-DNA interaction, while lipid-base binding and the stability of cationic lipid-DNA complexes have been the subject of more debate in recent years. The aim of this study was to examine the complexation of calf-thymus DNA with cholesterol (Chol), 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), dioctadecyldimethylammoniumbromide (DDAB) and dioleoylphosphatidylethanolamine (DOPE), at physiological condition, using constant DNA concentration and various lipid contents. Fourier transform infrared (FTIR), UV-visible, circular dichroism spectroscopic methods and atomic force microscopy were used to analyse lipid-binding site, the binding constant and the effects of lipid interaction on DNA stability and conformation. Structural analysis showed a strong lipid-DNA interaction via major and minor grooves and the backbone phosphate group with overall binding constants of KChol = 1.4 (±0.5) × 104M-1, KDDAB = 2.4 (±0.80) × 104M-1, KDOTAP = 3.1 (±0.90) × 104 M1 and KDOPE = 1.45 (±0.60) × 104M1. The order of stability of lipid-DNA complexation is DOTAP>DDAB>DOPE>Chol. Hydrophobic interactions between lipid aliphatic tails and DNA were observed. Chol and DOPE induced a partial B to A-DNA conformational transition, while a partial B to C-DNA alteration occurred for DDAB and DOTAP at high lipid concentrations. DNA aggregation was observed at high lipid content.
AB - Complexes of cationic liposomes with DNA are promising tools to deliver genetic information into cells for gene therapy and vaccines. Electrostatic interaction is thought to be the major force in lipid-DNA interaction, while lipid-base binding and the stability of cationic lipid-DNA complexes have been the subject of more debate in recent years. The aim of this study was to examine the complexation of calf-thymus DNA with cholesterol (Chol), 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), dioctadecyldimethylammoniumbromide (DDAB) and dioleoylphosphatidylethanolamine (DOPE), at physiological condition, using constant DNA concentration and various lipid contents. Fourier transform infrared (FTIR), UV-visible, circular dichroism spectroscopic methods and atomic force microscopy were used to analyse lipid-binding site, the binding constant and the effects of lipid interaction on DNA stability and conformation. Structural analysis showed a strong lipid-DNA interaction via major and minor grooves and the backbone phosphate group with overall binding constants of KChol = 1.4 (±0.5) × 104M-1, KDDAB = 2.4 (±0.80) × 104M-1, KDOTAP = 3.1 (±0.90) × 104 M1 and KDOPE = 1.45 (±0.60) × 104M1. The order of stability of lipid-DNA complexation is DOTAP>DDAB>DOPE>Chol. Hydrophobic interactions between lipid aliphatic tails and DNA were observed. Chol and DOPE induced a partial B to A-DNA conformational transition, while a partial B to C-DNA alteration occurred for DDAB and DOTAP at high lipid concentrations. DNA aggregation was observed at high lipid content.
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U2 - 10.1093/nar/gkn1003
DO - 10.1093/nar/gkn1003
M3 - Article
C2 - 19103664
AN - SCOPUS:63349088259
SN - 0305-1048
VL - 37
SP - 849
EP - 857
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 3
ER -