Phase Diagram, Stability, and Overcharging of Lamellar Cationic Lipid–DNA Self-Assembled Complexes

• Published in: Biophysical journal Vol. 77; no. 2; pp. 915 - 924
• Main Authors: Koltover, I., Salditt, T., Safinya, C.R.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.08.1999; Biophysical Society
• Subjects: Biophysical Phenomena; Biophysics; Cations; Deoxyribonucleic acid; DNA; DNA - administration & dosage; DNA - chemistry; DNA - genetics; Drug Carriers; Drug Stability; Gene Transfer Techniques; Lipids; Liposomes - chemistry; Macromolecular Substances; Models, Molecular; Molecular biology; Static Electricity
• DOI: 10.1016/S0006-3495(99)76942-0
• ISSN: 0006-3495

Cationic lipid–DNA (CL-DNA) complexes comprise a promising new class of synthetic nonviral gene delivery systems. When positively charged, they attach to the anionic cell surface and transfer DNA into the cell cytoplasm. We report a comprehensive x-ray diffraction study of the lamellar CL-DNA self-assemblies as a function of lipid composition and lipid/DNA ratio, aimed at elucidating the interactions determining their structure, charge, and thermodynamic stability. The driving force for the formation of charge-neutral complexes is the release of DNA and lipid counterions. Negatively charged complexes have a higher DNA packing density than isoelectric complexes, whereas positively charged ones have a lower packing density. This indicates that the overcharging of the complex away from its isoelectric point is caused by changes of the bulk structure with absorption of excess DNA or cationic lipid. The degree of overcharging is dependent on the membrane charge density, which is controlled by the ratio of neutral to cationic lipid in the bilayers. Importantly, overcharged complexes are observed to move toward their isoelectric charge-neutral point at higher concentration of salt co-ions, with positively overcharged complexes expelling cationic lipid and negatively overcharged complexes expelling DNA. Our observations should apply universally to the formation and structure of self-assemblies between oppositely charged macromolecules.