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Histone H1-mediated transfection: Serum inhibition can be overcome by Ca2+ ions

Item Type:Article
Title:Histone H1-mediated transfection: Serum inhibition can be overcome by Ca2+ ions
Creators Name:Haberland, A. and Knaus, T. and Zaitsev, S.V. and Buchberger, B. and Lun, A. and Haller, H. and Boettger, M.
Abstract:Purpose. One of the drawbacks of polycationic and cationic liposomal gene transfer is its sensitivity to serum. Gene therapy requires the transfectant-DNA complex to be resistant to serum as well as blood. Since Ca2+ has proved to be an efficient cofactor of polycationic gene transfer, we decided to investigate its effects on transfection in the presence of serum. Methods. We studied transgene expression of luciferase gene (pCMV Luc) on ECV 304 human endothelial cells using H1 histone and DOSPER as transfectants in the presence of 0-100% fetal calf serum. Results. H1-and DOSPER-mediated transfection was found to be inhibited by serum above the concentration of 10%. If 2 mM Ca2+ or 2 mM Ca2+/0.1 mM chloroquine was included in the culture medium which replace the transfection mixture and was left on the cells for 24 hours postincubation, the inhibiting effect of even 100% serum was overcome. Conclusions. A high serum level does not interfere with binding and uptake of H1- and DOSPER-DNA complexes, but inhibits subsequent steps such as endosomal escape. Ca2+ in the form of nascent calcium phosphate microprecipitates and other lysosomolytical agents facilitate endosomal/lysosomal release by their fusigenic and membranolytic activity.
Keywords:Calcium, DOSPER, HI Histone, Serum Inhibition, Transfection
Source:Pharmaceutical Research
ISSN:0724-8741
Publisher:Kluwer Academic/Plenum Publ (Germany)
Volume:17
Number:2
Page Range:229-235
Date:1 February 2000
Official Publication:https://doi.org/10.1023/A:1007581700996
PubMed:View item in PubMed

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