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Transgene expression is increased by photochemically mediated transduction of polycation-complexed adenoviruses

Gene Therapy volume 11, pages 152–160 (2004)Cite this article

Abstract

Poor efficiency of adenoviral gene transfer to target cells is a major limitation to adenoviral gene therapy. Inefficient gene transfer occurs in the absence of coxsackie- and adenovirus receptor (CAR) on the cell surface, and can be overcome by enhancing viral entry with cationic molecules. Recombinant adenovirus (Ad) noncovalently complexed with polycations imply a lack of transduction specificity. Therefore, we have investigated the potential of a novel light-specific treatment, named photochemical internalization (PCI), to enhance gene delivery of adenovirus serotype 5 (Ad5) complexed with the cationic agents poly-L-lysine (PLL) and SuperFect™. Cell lines differing in their receptiveness to Ad5 were infected with amounts of virus transducing about 2% of the cells by conventional Ad infection. The combination of polycations and photochemical treatment enabled a substantial increase in reporter gene expression, resulting in up to 75% positive cells. The effect was most prominent in cell lines expressing moderate to low levels of CAR. Furthermore, we show that PCI enables proper gene delivery of fiberless Ad5 at viral concentrations and infection times where transduction of photochemically untreated cells was negligible, both in the absence and presence of PLL. Thus, we conclude that the photochemically induced transduction by adenoviral vectors complexed with polycations present an opportunity to obtain high cell-infectivity levels with low viral doses, also without the fiber–CAR interaction.

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Acknowledgements

We are very grateful to Dr Frank L Graham for providing the Ad5CMV-lacZ adenovirus, and to Dr Dan von Seggern for providing the fiberless virus Ad5CMV-gfp-Δf. The work was supported by the Norwegian Research Council, the Norwegian Cancer Society and the Norwegian Ministry of Health.

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Authors and Affiliations

  1. Department of Biophysics, Institute for Cancer Research, The Norwegian Radium Hospital, Montebello, Norway

    A Bonsted, L Prasmickaite, O Kaalhus & K Berg

  2. Department of Tumor Biology, Institute for Cancer Research, The Norwegian Radium Hospital, Montebello, Oslo, Norway

    B Ø Engesæter & GM Mælandsmo

  3. PCI Biotech AS, Hoffsvn.48, Oslo, Norway

    A Høgset

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  1. A Bonsted
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Bonsted, A., Engesæter, B., Høgset, A. et al. Transgene expression is increased by photochemically mediated transduction of polycation-complexed adenoviruses. Gene Ther 11, 152–160 (2004). https://doi.org/10.1038/sj.gt.3302166

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