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. 2010 May 19;21(5):988-93.
doi: 10.1021/bc9005464.

The effect of charge-reversal amphiphile spacer composition on DNA and siRNA delivery

Xiao-Xiang Zhang  1 , Carla A H Prata, Thomas J McIntosh, Philippe Barthélémy, Mark W Grinstaff
Affiliations

The effect of charge-reversal amphiphile spacer composition on DNA and siRNA delivery

Xiao-Xiang Zhang et al. Bioconjug Chem. .

Abstract

A series of charge-reversal amphiphiles with different spacers separating the headgroup from the hydrophobic chains are described for delivery of DNA and siRNA. Among them, the amphiphiles possessing a glycine spacer (e.g., B-GlyGly) showed effective DNA transfection in CHO and NIH 3T3 cells, as well as siRNA gene knockdown in HepG2 and UASMC cells. Ethidium bromide quenching assays revealed that DNA was released the fastest from the lipoplex of B-GlyGly in the presence of esterase. Also, X-ray diffraction results indicated that the DNA was located between the adjacent lipid bilayers in the lipoplex of B-GlyGly. These distinct features appear to be required for high transfection activity.

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Figures

Figure 1
Figure 1
Amphiphiles previously studied.
Figure 2
Figure 2
Amphiphiles with different spacers.
Figure 3
Figure 3
EtBr displacement assay showing the fluorescence intensity. Top: as a function of amphiphile/DNA charge ratio; bottom: as a function of time in presence of porcine liver esterase (300 units/mL).
Figure 4
Figure 4
Sizes of lipoplexes (nm) at pH 7.4 (top) and pH 5 (bottom). N=3, mean±SD
Figure 5
Figure 5
DNA transfection after 48 h in CHO (top) and NIH3T3 (bottom) cells as a function of amphiphiles and DNA molar ratio. Lipofectamine™ 2000 was used as positive control. N=3, mean±SD. * p<0.05
Figure 6
Figure 6
Structures of B-Gly-C16 and B-GlyGly-C16 and DNA transfection after 48 h in CHO cells as a function of amphiphiles and DNA molar ratio. Lipofectamine™ 2000 was used as positive control. N=3, mean±SD
Figure 7
Figure 7
siRNA transfection after 48 h in HepG2 cells (top) and UASMC cells (bottom) as a function of amphiphiles and siRNA molar ratio, NeoFX™ was used as positive control. N=3, mean±SD
Scheme 1
Scheme 1
Synthesis of the cationic amphiphiles.
See this image and copyright information in PMC

References

    1. Mintzer MA, Simanek EE. Nonviral vectors for gene delivery. Chem Rev. 2009;109:259–302. - PubMed
    1. Ewert KK, Ahmad A, Bouxsein NF, Evans HM, Safinya CR. Non-viral gene delivery with cationic liposome-DNA complexes. Meth Mol Biol. 2008;433:159–175. - PubMed
    1. Christine CC, Huang L. Recent advances in non-viral gene delivery. Adv Genetics. 2005;53:3–18. - PubMed
    1. Wolff JA, Rozema DB. Breaking the bonds: Non-viral vectors become chemically dynamic. Mol Therapy. 2008;16:8–15. - PubMed
    1. Reineke TM, Grinstaff MW. Designer materials for nucleic acid delivery. Mat Res Soc Bull. 2005;30:635–639.

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