Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Viral Transfer Technology
  • Published:

Influence of promoter and WHV post-transcriptional regulatory element on AAV-mediated transgene expression in the rat brain

Gene Therapy volume 7, pages 1304–1311 (2000)Cite this article

Abstract

Recombinant adeno-associated viruses (rAAVs) can transduce several tissues, including the brain. However, in brain the duration of gene expression in different areas is variable, which has been ascribed to viral (CMV) promoter silencing in some regions over time. We have compared expression of enhanced green fluorescent protein (EGFP) in the nigrostriatal pathway of rats mediated by rAAVs containing the CMV or platelet-derived growth factor-β chain (PDGF-β) promoter. In addition, we studied the effects of the woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) on transgene expression in vivo. The rAAV vectors containing the neuron-specific PDGF-β chain promoter transduced significantly more dopaminergic neurons than titer-matched vectors carrying the CMV promoter. Moreover, the WPRE further increased EGFP expression, and a rAAV vector incorporating both the PDGF-β chain promoter and the WPRE resulted in efficient EGFP expression in dopaminergic neurons and their projections in the striatum for at least 41 weeks after virus injection. Our results emphasize the importance of a strong tissue-specific promoter in achieving optimal transgene expression, not only in long-term but also in short-term studies where viral titers may be limiting. Furthermore, they suggest that incorporation of the WPRE into rAAVs, and possibly other types of vectors, is useful to enhance transgene expression in vivo.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Rabinowitz JE, Samulski J . Adeno-associated virus expression systems for gene transfer Curr Opin Biotechnol 1998 9: 470–475

    Article  CAS  PubMed  Google Scholar 

  2. Büeler H . Adeno-associated viral vectors for gene transfer and gene therapy Biol Chem 1999 380: 613–622

    Article  PubMed  Google Scholar 

  3. Xiao X, Li J, Samulski RJ . Efficient long-term gene transfer into muscle tissue of immunocompetent mice by adeno-associated virus vector J Virol 1996 70: 8098–8108

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Flannery JG et al. Efficient photoreceptor-targeted gene expression in vivo by recombinant adeno-associated virus Proc Natl Acad Sci USA 1997 94: 6916–6921

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Mandel RJ, Spratt SK, Snyder RO, Leff SE . Midbrain injection of recombinant adeno-associated virus encoding rat glial cell line-derived neurotrophic factor protects nigral neurons in a progressive 6-hydroxydopamine-induced degeneration model of Parkinson's disease in rats Proc Natl Acad Sci USA 1997 94: 14083–14088

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Snyder RO et al. Persistent and therapeutic concentrations of human factor IX in mice after hepatic gene transfer of recombinant AAV vectors Nat Genet 1997 16: 270–276

    Article  CAS  PubMed  Google Scholar 

  7. During MJ et al. In vivo expression of therapeutic human genes for dopamine production in the caudates of MPTP-treated monkeys using an AAV vector Gene Therapy 1998 5: 820–827

    Article  CAS  PubMed  Google Scholar 

  8. Herzog RW et al. Long-term correction of canine hemophilia B by gene transfer of blood coagulation factor IX mediated by adeno-associated viral vector Nature Med 1999 5: 56–63

    Article  CAS  PubMed  Google Scholar 

  9. Jooss K, Yang Y, Fisher KJ, Wilson JM . Transduction of dendritic cells by DNA viral vectors directs the immune response to transgene products in muscle fibers J Virol 1998 72: 4212–4223

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Brockstedt DG, Podsakoff GM, Fong L, Kurtzman G, Mueller-Ruchholtz W, Engleman EG . Induction of immunity to antigens expressed by recombinant adeno-associated virus depends on the route of administration Clin Immunol 1999 92: 67–75

    Article  CAS  PubMed  Google Scholar 

  11. Bartlett JS, Samulski RJ, McCown TJ . Selective and rapid uptake of adeno-associated virus type 2 in brain Hum Gene Ther 1998 9: 1181–1186

    Article  CAS  PubMed  Google Scholar 

  12. Mandel RJ et al. Characterization of intrastriatal recombinant adeno-associated virus-mediated gene transfer of human tyrosine hydroxylase and human GTP-cyclohydrolase I in a rat model of Parkinson's disease J Neurosci 1998 18: 4271–4284

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. McCown TJ et al. Differential and persistent expression patterns of CNS gene transfer by an adeno-associated virus (AAV) vector Brain Res 1996 713: 99–107

    Article  CAS  PubMed  Google Scholar 

  14. Klein RL et al. Neuron-specific transduction in the rat septohippocampal or nigrostriatal pathway by recombinant adeno-associated virus vectors Exp Neurol 1998 150: 183–194

    Article  CAS  PubMed  Google Scholar 

  15. Klein RL et al. Long-term actions of vector-derived nerve growth factor or brain-derived neurotrophic factor on choline acetyltransferase and Trk receptor levels in the adult rat basal forebrain Neuroscience 1999 90: 815–821

    Article  CAS  PubMed  Google Scholar 

  16. Donello JE, Loeb JE, Hope TJ . Woodchuck hepatitis virus contains a tripartite posttranscriptional regulatory element J Virol 1998 72: 5085–5092

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Zufferey R, Donello JE, Trono D, Hope TJ . Woodchuck hepatitis virus posttranscriptional regulatory element enhances expression of transgenes delivered by retroviral vectors J Virol 1999 73: 2886–2892

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Loeb JE et al. Enhanced expression of transgenes from adeno-associated virus vectors with the woodchuck hepatitis virus posttranscriptional regulatory element: implications for gene therapy Hum Gene Ther 1999 10: 2295–2305

    Article  CAS  PubMed  Google Scholar 

  19. Sasahara M et al. PDGF B-chain in neurons of the central nervous system, posterior pituitary, and in a transgenic model Cell 1991 64: 217–227

    Article  CAS  PubMed  Google Scholar 

  20. Douglas R, Kellaway L, Mintz M, van Wageningen G . The crossed nigrostriatal projection decussates in the ventral tegmental decussation Brain Res 1987 418: 111–121

    Article  CAS  PubMed  Google Scholar 

  21. Xiao X, Li J, McCown TJ, Samulski RJ . Gene transfer by adeno-associated virus vectors into the central nervous system Exp Neurol 1997 144: 113–124

    Article  CAS  PubMed  Google Scholar 

  22. Szczypka MS et al. Viral gene delivery selectively restores feeding and prevents lethality of dopamine-deficient mice Neuron 1999 22: 167–178

    Article  CAS  PubMed  Google Scholar 

  23. Kaplitt MG et al. Long-term gene expression and phenotypic correction using adeno-associated virus vectors in the mammalian brain Nat Genet 1994 8: 148–154

    Article  CAS  PubMed  Google Scholar 

  24. Prosch S et al. Inactivation of the very strong HCMV immediate early promoter by DNA CpG methylation in vitro Biol Chem 1996 377: 195–201

    CAS  Google Scholar 

  25. Loser P, Jennings GS, Strauss M, Sandig V . Reactivation of the previously silenced cytomegalovirus major immediate–early promoter in the mouse liver: involvement of NFkappaB J Virol 1998 72: 180–190

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Games D et al. Alzheimer-type neuropathology in transgenic mice overexpressing V717F beta-amyloid precursor protein Nature 1995 373: 523–527

    Article  CAS  PubMed  Google Scholar 

  27. Peel AL et al. Efficient transduction of green fluorescent protein in spinal cord neurons using adeno-associated virus vectors containing cell type-specific promoters Gene Therapy 1997 4: 16–24

    Article  CAS  PubMed  Google Scholar 

  28. Samulski RJ, Chang LS, Shenk T . Helper-free stocks of recombinant adeno-associated viruses: normal integration does not require viral gene expression J Virol 1989 63: 3822–3828

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Snyder RO, Xiao X, Samulski RJ . Production of recombinant adeno-associated viral vectors. In: Dracopoli NC et al (eds) Current Protocols in Human Genetics John Wiley & Sons 1997 pp . 12.1.1–12.1.20.

Download references

Acknowledgements

We thank S Furler for counting neurons and acknowledge I Allmann for providing rats. We are grateful to S Hemmi and Z Dong for critically reading the manuscript, to F Ochsenbein for preparing figures and to A Gentilini and L Bordoli for help with statistical analysis. This work was supported by grants of the Swiss National Science Foundation 31-044449.95 and 31-046648.96 to HB.

Author information

Authors and Affiliations

  1. Institute of Molecular Biology, University of Zurich, Zurich, Switzerland

    J-C Paterna, T Moccetti & H Büeler

  2. Institute of Toxicology, Swiss Federal Institute of Technology, Zurich, Switzerland

    A Mura & J Feldon

Authors
  1. J-C Paterna
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T Moccetti
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A Mura
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J Feldon
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H Büeler
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Paterna, JC., Moccetti, T., Mura, A. et al. Influence of promoter and WHV post-transcriptional regulatory element on AAV-mediated transgene expression in the rat brain. Gene Ther 7, 1304–1311 (2000). https://doi.org/10.1038/sj.gt.3301221

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301221

Keywords

This article is cited by

Search

Advanced search

Quick links