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  • Viral Transfer Technology
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Sustained ex vivo and in vivo transfer of a reporter gene using polyoma virus pseudocapsids

Gene Therapy volume 7, pages 1094–1102 (2000)Cite this article

Abstract

Properties of a virus-like artificial gene delivery vehicle, synthesised from recombinant major coat protein of mouse polyoma virus, have been explored. The protein, VP1, self assembles into protein spheres, or ‘pseudocapsids’, which can bind and transfer DNA into cells in vitro and in vivo. Here, the ability of pseudocapsids to carry DNA into a complex cell system (ex vivo organ cultures of rabbit cornea) or whole animals (mice) has been assessed. Evidence from histochemical and PCR experiments indicate that pseudocapsids stimulate uptake and stable maintenance of marker DNA in nondividing corneal cells as efficiently as a recombinant adenovirus. In athymic and immunocompetent mice, gene transmission occurs with no apparent adverse effects on the animals. In the presence of pseudocapsids, the marker gene was transferred to a range of organs, including the brains of animals, following peripheral or intranasal administration. In immunocompetent mice, significant long-term transcriptional expression (at least 22 weeks) was observed with pseudocapsids, a period significantly longer than observed with DNA alone (several weeks only), again with no obvious adverse effects. This study demonstrates that pseudocapsids from the murine virus, polyoma, constitute a novel transfer agent for long-term gene therapeutic applications in tissues or whole animals.

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References

  1. Verma IM, Somia N . Gene therapy – promises, problems and prospects Nature 1997 389: 239–242

    Article  CAS  Google Scholar 

  2. Anderson WF . Human gene therapy Nature 1998 392: 25–30

    CAS  Google Scholar 

  3. Montross L et al. Nuclear assembly of polyomavirus capsids in insect cells expressing the major capsid protein VP1 J Virol 1991 65: 4991–4998

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Forstová J et al. Cooperation of structural proteins during late events in the life cycle of polyomavirus J Virol 1993 67: 1405–1413

    PubMed  PubMed Central  Google Scholar 

  5. Rodgers RE, Chang D, Cai X, Consigli RA . Purification of recombinant budgerigar fledgling disease virus VP1 capsid protein and its ability for in vitro capsid assembly J Virol 1994 68: 3386–3390

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Chang D et al. Self-assembly of the JC virus major capsid protein, VP1, expressed in insect cells J Gen Virol 1997 78: 1435–1439

    Article  CAS  Google Scholar 

  7. Pawlita M et al. DNA encapsidation by virus like particles assembled in insect cells from the major capsid protein VP1 of B-lymphotropic papovavirus J Virol 1996 70: 7517–7526

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Shishido Y et al. Assembly of JC virus-like particles in COS7 cells J Med Virol 1997 51: 265–272

    Article  CAS  Google Scholar 

  9. Sandalon Z, Oppenheim A . Self-assembly and protein–protein interactions between the SV40 capsid proteins produced in insect cells Virology 1997 237: 414–421

    Article  CAS  Google Scholar 

  10. Touze A et al. Production of recombinant virus-like particles from human papillomavirus types 6 and 11, and study of serological reactivities between HPV 6, 11, 16 and 45 by ELISA: implications for papillomavirus prevention and detection Fems Microbiol Lett 1998 160: 111–118

    Article  CAS  Google Scholar 

  11. Touze A et al. Production of human papillomavirus type 45 virus-like particles in insect cells using a recombinant baculovirus Fems Microbiol Lett 1996 141: 111–116

    Article  CAS  Google Scholar 

  12. Garcea RL, Liddington RC . Structural biology of polyomaviruses. In: Chiu W, Burnett RM, Garcea RL (eds) Structural Biology of Viruses Oxford University Press: Oxford 1997 187–208

    Google Scholar 

  13. Forstová J et al. Polyoma virus pseudocapsids as efficient carriers of heterologous DNA into mammalian cells Hum Gene Ther 1995 6: 297–306

    Article  Google Scholar 

  14. Touze A, Coursaget P . In vitro gene transfer using human papillomavirus-like particles Nucleic Acids Res 1998 26: 1317–1323

    Article  CAS  Google Scholar 

  15. Strayer DS, Milano J . SV40 mediates stable gene transfer in vivo Gene Therapy 1996 3: 581–587

    CAS  Google Scholar 

  16. Strayer DS . SV40 as an effective gene transfer vector in vivo J Biol Chem 1996 271: 24741–24746

    CAS  Google Scholar 

  17. Strayer DS, Kondo R, Milano J, Duan LX . Use of SV40-based vectors to transduce foreign genes to normal human peripheral blood mononuclear cells Gene Therapy 1997 4: 219–225

    Article  CAS  Google Scholar 

  18. Oppenheim A, Peleg A, Fibach E, Rachmilewitz EA . Efficient introduction of plasmid DNA into human hemopoietic cells by encapsidation in simian virus 40 pseudovirions Proc Natl Acad Sci USA 1986 83: 6925–6929

    Article  CAS  Google Scholar 

  19. Rund D et al. Efficient transduction of human hematopoietic cells with the human multidrug resistance gene 1 via SV40 pseudovirions Hum Gene Ther 1998 9: 649–657

    Article  CAS  Google Scholar 

  20. Soeda E et al. Enhancement by polylysine of transient, but not stable, expression of genes carried into cells by polyoma VP1 pseudocapsids Gene Therapy 1998 5: 1410–1419

    Article  CAS  Google Scholar 

  21. Soeda E . Developing polyoma viral pseudocapsids for gene transfer; delivery into cells with poly-l-lysine PhD thesis, University of London 1999

    Google Scholar 

  22. Bondi A et al. The use of beta-galactosidase as a tracer in immunocytochemistry Histochemistry 1982 76: 153–158

    Article  CAS  Google Scholar 

  23. Gluzman Y . SV40-transformed simian cells support the replication of early SV40 mutants Cell 1981 23: 175–182

    Article  CAS  Google Scholar 

  24. Larkin DFP et al. Adenovirus-mediated gene delivery to the corneal epithelium Transplantation 1996 61: 363–370

    Article  CAS  Google Scholar 

  25. Wilkinson WGW, Akrigg A . Constitutive expression from the CMV major IE promoter in a defective adenovirus Nucleic Acids Res 1992 20: 2233–2239

    Article  CAS  Google Scholar 

  26. Sun N, Cassell MD, Perlman S . Anterograde, transneuronal transport of herpes simplex virus type 1 strain H129 in the murine visual system J Virol 1996 70: 5405–5413

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Engel JP, Madigan TC, Peterson GM . The transneuronal spread phenotype of herpes simplex virus type 1 infection of the mouse hind footpad J Virol 1997 71: 2425–2435

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Draghia R et al. Gene delivery into the central nervous system by nasal instillation in rats Gene Therapy 1995 2: 418–423

    CAS  PubMed  Google Scholar 

  29. Berke Z et al. Persistence of polyomavirus in adult SCID C.B-17 mice In Vivo 1994 8: 339–342

    CAS  PubMed  Google Scholar 

  30. Sebesteny A, Tilly R, Balkwill F, Trevan D . Demyelination and wasting associated with polyomavirus infection in nude (nu/nu) mice Lab Anim 1980 14: 337–345

    Article  CAS  Google Scholar 

  31. McCance DJ et al. A paralytic disease in nude mice associated with polyoma virus infection J Gen Virol 1983 64: 57–67

    Article  Google Scholar 

  32. Elsner C, Dorries K . Evidence of human polyomavirus BK and JC infection in normal brain tissue Virology 1992 191: 72–80

    Article  CAS  Google Scholar 

  33. Rowe WP, Huebner RJ, Hartley JW . Ecology of a mouse tumour virus. In: Pollard M (ed) Perspectives in Virology II Burgess Publishing Co: Minneapolis 1961 177–190

    Google Scholar 

  34. Pontén J . Spontaneous and Virus-induced Transformation in Cell Culture Springer-Verlag: Vienna, New York 1971 pp 51–56

    Chapter  Google Scholar 

  35. Bronson R, Dawe C, Carroll J, Benjamin T . Tumor induction by a transformation-defective polyoma virus mutant blocked in signaling through Shc Proc Natl Acad Sci USA 1997 94: 7954–7958

    Article  CAS  Google Scholar 

  36. Salunke DM, Caspar DL, Garcea RL . Self-assembly of purified polyomavirus capsid protein VP1 Cell 1986 46: 895–904

    Article  CAS  Google Scholar 

  37. Ou W-C et al. The major capsid protein, VP1, of human JC virus expressed in Escherichia coli is able to self-assemble into a capsid-like particle and deliver exogenous DNA into human kidney cells J Gen Virol 1999 80: 39–46

    Article  CAS  Google Scholar 

  38. Goldmann C et al. Molecular cloning and expression of major structural protein VP1 of the human polyomavirus JC virus: formation of virus-like particles useful for immunological and therapeutic studies J Virol 1999 73: 4465–4469

    CAS  PubMed  PubMed Central  Google Scholar 

  39. Cox C et al. Persistent Epstein–Barr virus infection in the common marmoset (Callithrix jacchus) J Gen Virol 1996 77: 1173–1180

    Article  CAS  Google Scholar 

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Acknowledgements

We thank K Harrington for assistance with the intravenous injections. This work was supported by The Wellcome Trust (award 048711/2/96/2), the Biotechnology and Biological Sciences Research Council (grant 60/4291) and the European Community (BIO4-CT97-2147). SR acknowledges support from the Iris Fund for the Prevention of Blindness.

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Author notes

  1. N Krauzewicz & B Clark

    Present address: MRC Clinical Sciences Centre, ICSM, Hammersmith Hospital, Du Cane Road, London, W12 ONN, UK

Authors and Affiliations

  1. Department of Infectious Diseases (Virology), Imperial College School of Medicine at Hammersmith Hospital, London, UK

    N Krauzewicz, C Cox, E Soeda & BE Griffin

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

    BE Griffin

  3. Department of Immunology, Imperial College School of Medicine at Hammersmith Hospital, London, UK

    S Rayner

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Krauzewicz, N., Cox, C., Soeda, E. et al. Sustained ex vivo and in vivo transfer of a reporter gene using polyoma virus pseudocapsids. Gene Ther 7, 1094–1102 (2000). https://doi.org/10.1038/sj.gt.3301219

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