caa a22 4500 477128300 CHVBK 20180405111656.0 cr unu---uuuuu 170330e19970201xx s 000 0 eng 10.1023/A:1012000711033 doi (NATIONALLICENCE)springer-10.1023/A:1012000711033 Self-Assembling DNA-Lipid Particles for Gene Transfer [Elektronische Daten] [Yuan-Peng Zhang, Dorothy Reimer, Guoyang Zhang, Patricia Lee, Marcel Bally] Purpose. We have demonstrated that a heteromolecular complex consisting of cationic lipids and DNA can be prepared and isolated (1). Cationic lipids bind DNA through electrostatic interactions. However, when sufficient lipids are bound to DNA the physical and chemical properties of the complex are governed by hydrophobic effects. Here we describe an approach where this hydrophobic complex is used as an intermediate in the preparation of lipid-DNA particles (LDPs). Methods. The approach relies on the generation of mixed micelles containing the detergent, n-octyl β-D-glucopyranoside (OGP), the cationic lipid, N-N-dioleoyl-N, N-dimethylammonium chloride (DODAC), and selected zwitterionic lipids, 1,2-dioleoyl-sn-glycero-3 -phosphoethanolamine (DOPE) or egg sphingomyelin (SM). Results. When these micelles were prepared at low detergent concentrations (20 mM OGP) and combined with pCMVβ DNA, LDPs spontaneously formed. The mean diameter of these particles as measured by quasielastic light scattering was 55−70 nm, a result that was confirmed by negative stain electron microscopy. Further characterization of these LDPs showed that DNA within the particles was inaccessible to the small fluorochrome TO-PRO-1 and protected against DNase I degradation. LDPs could also be prepared in high concentrations of OGP (100 mM), however particles formed only after removal of OGP by dialysis. Particles formed in this manner were large (>2000nm) and mediated efficient transfection of Chinese hamster ovary cells. Transfection activity was greater when the lipid composition used consisted of SM/ DODAC. Small particles (<100nm) prepared of SM/DODAC were, however, inefficient transfecting agents. Conclusions. We believe that LDP formation is a consequence of the molecular forces that promote optimal hydrocarbon-hydrocarbon interactions and elimination of the hydrocarbon-water interface. Plenum Publishing Corporation, 1997 gene transfer nationallicence cationic lipid nationallicence DNA complexes nationallicence hydrophobic effect nationallicence Zhang Yuan-Peng aut Reimer Dorothy aut Zhang Guoyang aut Lee Patricia aut Bally Marcel aut Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/2(1997-02-01), 190-196 0724-8741 14:2<190 1997 14 11095 https://doi.org/10.1023/A:1012000711033 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1012000711033 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zhang Yuan-Peng aut NATIONALLICENCE 700 1- Reimer Dorothy aut NATIONALLICENCE 700 1- Zhang Guoyang aut NATIONALLICENCE 700 1- Lee Patricia aut NATIONALLICENCE 700 1- Bally Marcel aut NATIONALLICENCE 773 0- Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/2(1997-02-01), 190-196 0724-8741 14:2<190 1997 14 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer