DNA condensation, structure and transfection efficiency of complexes formed by gemini surfactants alkane-α,ω-diyl-bis(dodecyldimethylammonium bromide)s (CnGS12, n = 3, 6 and 12 is the number of alkane spacer carbons), dioleoylphosphatidylethanolamine (CnGS12/DOPE = 0.3 mol/ mol) and DNA at low surface charge density were investigated through different techniques. Small angle X-ray diffraction showed a condensed lamellar phase with marked dependence of DNA-DNA distance on (+/-) charge ratio. High ionic strength of hydrating medium screens the interaction DNA -CnGS12/DOPE and complexed DNA represented maximally ~ 45-60% of total DNA in the solution as derived from fluorescence and UV-VIS spectroscopy. The in vitro transfection efficiency of CnGS12/DOPE liposomes on mammalian HEK 293 cell line was spacer length-dependent. C12GS12/DOPE/DNA complexes exhibited the best transfection efficiency (~ 18% GFP-expressing cells relative to all viable cells) accompanied by ~ 89% cell viability.
Small angle X-ray diffractionFigure S1. SAXD patterns of C6GS12/DOPE/DNA lipoplexes at molar ratio C6GS12/DOPE = 0.3 and charge ratios C6GS12/DNA = 1-3 hydrated by mixture of Opti-MEM and DMEM medium. The relative intensity is in logarithmic scale. Doi: 10.4149/gpb_2017042 S u p p l e m e n t a r y M a t e r i a l DNA-DOPE-gemini surfactants complexes at low surface charge density: from structure to transfection efficiency Small angle X-ray diffraction Figure S1. SAXD patterns of C6GS12/DOPE/DNA lipoplexes at molar ratio C6GS12/DOPE = 0.3 and charge ratios C6GS12/DNA = 1-3 hydrated by mixture of Opti-MEM and DMEM medium. The relative intensity is in logarithmic scale.
Phospholipid-based non-viral carriers composed of neutral phospholipid dioleoylphosphatidylethanolamine (DOPE) and the binary mixture DOPE–oleic acid (OA) are examined as potential DNA delivery vectors. The process of DNA condensation in the presence of Ca2+ ions has been monitored through changes in emmision intensity of fluorescent probe ethidium bromide. The decline in fluorescence intensity with increasing Ca2+ concentration at two different time intervals was correlated with the binding capacity of complexes and possible release of DNA from the complex. The microstructure of DOPE–OA mixtures at different OA/DOPE molar ratios and that of DOPE–OA–DNA–Ca2+ complexes were determined using synchrotron small angle X-ray diffraction (SAXD). We identified inverted hexagonal phase HII as the dominant structure. OA affects the lattice parameter of HII formed by DOPE. With the increasing OA/DOPE molar ratio, the lattice parameter decreases, which results in significantly lower fraction of DNA bound to the OA-enriched complexes.
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