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Issue 2, 2011
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Lipid chain geometry of C14 glycerol-based lipids: effect on lipoplex structure and transfection

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The effects have been determined of a systematic alteration of the alkyl chain geometry of a C14 analogue of DOTMA on the detailed molecular architecture of the resulting cationic vesicles formed both in the absence and presence of 50 mol% DOPE, and of the lipoplexes prepared from these vesicles using either calf thymus or plasmid DNA. The C14 DOTMA analogues studied involved cis- or trans-double bonds at positions Δ9 or Δ11, and a compound (ALK) featuring an alkyne at position C9. For all of these analogues, examination by light scattering and neutron scattering, zeta potential measurement, and negative staining electron microscopy showed that there were no significant differences in the structures or charges of the vesicles or of the resulting lipoplexes, regardless of the nature of the DNA incorporated. Differences were observed, however, between the complexes formed by the various lipids when examining the extent of complexation and release by gel electrophoresis, where the E-lipids appeared to complex the DNA more efficiently than all other lipids tested. Moreover, the lipoplexes prepared from the E-lipids were the most effective in transfection of MDA-MB-231 breast cancer cells. As indicated through confocal microscopy studies, the E-lipids also showed a higher internalisation capacity and a more diffuse cellular distribution, possibly indicating a greater degree of endosomal escape and/or nuclear import. These observations suggest that the extent of complexation is the most important factor in determining the transfection efficiency of the complexes tested. At present it is unclear why the E-lipids were more effective at complexing DNA, although it is thought that the effective area per molecule occupied by the cationic lipid and DOPE head groups, and therefore the density of positive charges on the surface of the bilayer most closely matches the negative charge density of the DNA molecule. From a consideration of the geometry of the cationic lipids it is anticipated that the head groups of the E-lipids would occupy a smaller area per molecule than the ALK or Z-lipids.

Graphical abstract: Lipid chain geometry of C14 glycerol-based lipids: effect on lipoplex structure and transfection

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Publication details

The article was received on 10 Aug 2010, accepted on 30 Sep 2010 and first published on 08 Nov 2010

Article type: Paper
DOI: 10.1039/C0MB00149J
Citation: Mol. BioSyst., 2011,7, 422-436
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    Lipid chain geometry of C14 glycerol-based lipids: effect on lipoplex structure and transfection

    L. Kudsiova, J. Ho, B. Fridrich, R. Harvey, M. Keppler, T. Ng, S. L. Hart, A. B. Tabor, H. C. Hailes and M. J. Lawrence*, Mol. BioSyst., 2011, 7, 422
    DOI: 10.1039/C0MB00149J

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