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  • Viral Transfer Technology
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Virus-like gene transfer into cells mediated by polyoma virus pseudocapsids

Gene Therapy volume 7, pages 2122–2131 (2000)Cite this article

Abstract

Mouse polyoma virus-like particles (or pseudocapsids) are composed solely of recombinant viral coat protein. They can interact with DNA and transport it to cells, resulting in gene expression both in tissue culture and in mice. We demonstrate that DNA transfer in vitro depends on partial packaging of DNA within the virus-like capsid. Cell surface sialic acid residues and an intact microtubule network, required for viral infectivity, are also necessary for pseudocapsid-mediated gene expression from heterologous DNA. Thus, gene delivery in this system requires pathways utilised by polyoma virions, rather than proceeding via the ‘nonspecific’ endosomal route typical of nonviral systems such as liposomes or calcium phosphate precipitates. Despite the fact that all cells appear to internalise pseudocapsid/DNA complexes, only a proportion show productive gene delivery. Bulk internalisation of complexes is dependent on actin fibres, but not cell surface sialic acid or microtubules, indicating that a second transport pathway exists for pseudocapsids which is nonproductive for gene transfer. The model suggested by these data demonstrates the virus-like properties of the pseudocapsid system, and provides a basis for further development to produce a highly effective gene delivery vehicle.

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Acknowledgements

We thank M Stevens and J Forstová for many helpful discussions, A Sardini and G Warnes for discussion and assistance with light microscopy and flow cytometry and I Robinson for help with AFM. Support is acknowledged from The Wellcome Trust (Grant No. 048711/Z/96), the Medical Research Council and the European Community (No. BIO4-CT97–2147). The scanning force microscope was purchased with assistance from the HEFCW Technology Foresight Initiative.

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Authors and Affiliations

  1. MRC Clinical Sciences Centre, Imperial College School of Medicine, Hammersmith Hospital, Ducane Road, London, W12 0NN, UK

    N Krauzewicz & C F Higgins

  2. Institute of Molecular Genetics, Czech Academy of Sciences, Prague, Czech Republic

    J Stokrová

  3. Department of Physics and Astronomy, University of Wales Cardiff, Cardiff

    C Jenkins & M Elliott

  4. Division of Medicine, Viral Oncology Unit, Imperial College School of Medicine at St Mary's, London, UK

    B E Griffin

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  1. N Krauzewicz
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Krauzewicz, N., Stokrová, J., Jenkins, C. et al. Virus-like gene transfer into cells mediated by polyoma virus pseudocapsids. Gene Ther 7, 2122–2131 (2000). https://doi.org/10.1038/sj.gt.3301322

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