Structural dynamics and physicochemical properties of pDNA/DODAB:MO lipoplexes: Effect of pH and anionic lipids in inverted non-lamellar phases versus lamellar phases
作者： J.P. Neves Silva , I.M.S.C. Oliveira , A.C.N. Oliveira , M. Lúcio , A.C. Gomes
DOI: 10.1016/J.BBAMEM.2014.06.014
关键词:
摘要: Dioctadecyldimethylammonium bromide (DODAB):Monoolein (MO) lipoplexes have mainly been studied within the range of high molar ratios DODAB, with noticeable transfection efficiencies in Human Embryonic Kidney (HEK, a.k.a. 293T) cell line. In this work, we intend to study effect MO content on structure and physicochemical properties pDNA/DODAB:MO achieve some correlation their efficiency. Static/Dynamic Light Scattering Cryo-TEM imaging were used characterize size/morphology DNA/DODAB:MO at different DODAB:MO contents (2:1, 1:1, 1:2) charge (CRs) (+/-). Nile Red fluorescence emission was performed detect changes microviscosity, hydration polarity systems. Lipoplexes stability physiological pH values presence anionic lipids evaluated by Forster Resonance Energy Transfer (FRET). Physicochemical/structural data complemented studies HEK cells using β-galactosidase reporter gene activity assay. This work reports coexistence multilamellar non-lamellar inverted phases MO-richer (DODAB:MO 1:2 1:4), leading comparable those (DODAB-richer) lipoplexes, but higher [CR (+/-)=6.0] without dose-effect response. These results may be related structural promoted content.
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