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.

  • Brief Communication
  • Published:

Insulation of a conditionally expressed transgene in an adenoviral vector

Abstract

Replication-defective recombinant adenoviruses provide an efficient system for in vivo gene transfer and numerous studies have demonstrated that this vector can accommodate tissue-specific promoters to restrict the expression of a transgene to a particular subset of cells. However, in some cases the selectivity of expression is lost when the tissue-specific promoter is placed in an adenoviral environment. In an attempt to restore the conditionality of expression of the transgene driven by the human ERBB2 promoter, we have flanked the expression cassette in 5′ and 3′ orientations with a 250 bp sequence containing the bovine growth hormone transcriptional stop signal for cloning into a recombinant adenovirus. The data presented here clearly demonstrate that these ‘insulator’ elements are able to restrict the expression of the transgene (herpes simplex thymidine kinase) to ERBB2-expressing cells and therefore to restore the selectivity mediated by the ERBB2 promoter. This approach could be generally useful to insulate expression cassettes in adenoviral vectors.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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. Douglas JT et al. Targeted gene delivery by tropism-modified adenoviral vectors Nature Biotech 1996 14: 1574–1578

    Article  CAS  Google Scholar 

  2. Gutierrez AA, Lemoine NR, Sikora K . Gene therapy for cancer Lancet 1992 339: 715–721

    Article  CAS  Google Scholar 

  3. Rogers BE et al. Enhanced in vivo gene delivery to human ovarian cancer xenografts utilising a tropism-modified adenovirus vector Tumor Targeting 1998 3: 25–31

    CAS  Google Scholar 

  4. Siders WM, Halloran PJ, Fenton RG . Transcriptional targeting of recombinant adenoviruses to human and murine melanoma cells Cancer Res 1996 56: 5638–5646

    CAS  PubMed  Google Scholar 

  5. Rothmann T et al. Heart muscle-specific gene expression using replication defective recombinant adenovirus Gene Therapy 1996 3: 919–926

    CAS  PubMed  Google Scholar 

  6. Larochelle N et al. Efficient muscle-specific transgene expression after adenovirus-mediated gene transfer in mice using a 1.35 kb muscle creatine kinase promoter/enhancer Gene Therapy 1997 4: 465–472

    Article  CAS  Google Scholar 

  7. Kim S et al. Transcriptional targeting of replication-defective adenovirus transgene expression to smooth muscle cells in vivo J Clin Invest 1997 100: 1006–1014

    Article  CAS  Google Scholar 

  8. Inesi G et al. Cell-specific promoter in adenovirus vector for transgenic expression of SERCA1 ATPase in cardiac myocytes Am J Physiol 1998 274: C645–C653

    Article  CAS  Google Scholar 

  9. Ring CJA, Harris JD, Hurst HC, Lemoine NR . Suicide gene expression in tumour cells transduced with recombinant adenoviral, retroviral and plasmid vectors containing the ERBB2 promoter Gene Therapy 1996 3: 1094–1103

    CAS  PubMed  Google Scholar 

  10. Shi Q, Wang Y, Worton D . Modulation of the specificity and activity of a cellular promoter in an adenoviral vector Hum Gene Ther 1997 8: 403–410

    Article  CAS  Google Scholar 

  11. Slamon DJ et al. Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene Science 1987 235: 177–182

    Article  CAS  Google Scholar 

  12. Hall PA et al. The c-erbB-2 proto-oncogene in human pancreatic cancer J Pathol 1990 161: 195–200

    Article  CAS  Google Scholar 

  13. Lemoine NR et al. Amplification and over-expression of the EGF receptor and c-erbB-2 oncogenes in human stomach cancer Br J Cancer 1991 64: 74–83

    Article  Google Scholar 

  14. Hollywood D, Hurst H . A novel transcription factor, OB2–1, is required for over-expression of the proto-oncogene c-erbB-2 in mammary tumour lines EMBO J 1993 12: 2369–2375

    Article  CAS  Google Scholar 

  15. Martin LA, Lemoine NR . Direct cell killing by suicides genes Cancer Metas Rev 1996 15: 301–316

    Article  CAS  Google Scholar 

  16. Mittereder N, March KL, Trapnell BC . Evaluation of the concentration and bioactivity of adenovirus vector for gene therapy J Virol 1996 70: 7498–7509

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Kalos M, Fournier RE . Position-independent transgene expression mediated by boundary elements from the apolipoprotein B chromatin domain Mol Cell Biol 1995 15: 198–207

    Article  CAS  Google Scholar 

  18. Wang DM, Taylor S, Levy-Wilson B . Evaluation of the function of the human apolipoprotein B gene matrix attachment regions in transgenic mice J Lipid Res 1996 37: 2117–2124

    CAS  PubMed  Google Scholar 

  19. Makarova O et al. Incorporation of nuclear matrix attachment regions into the herpes simplex virus type 1 genome does not induce long-term expression of a foreign gene during latency Gene Therapy 1996 3: 829–833

    CAS  PubMed  Google Scholar 

  20. Pikaart MJ, Recillas-Targa F, Felsenfeld G . Loss of transcriptional activity of a transgene is accompanied by DNA methylation and histone deacetylation and is prevented by insulators Gene Dev 1998 12: 2852–2862

    Article  CAS  Google Scholar 

  21. Bett AJ, Krougliak V, Graham FL . DNA sequence of the deletion/insertion in early region 3 of Ad5 dl309 Virus Res 1995 39: 75–82

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We would like to thank Steve Murphy for advice in the preparation of the recombinant Ad5-Ins-1ETAS and Lesley-Ann Martin and Anne Rigg for critical reading of the manuscript. This work was supported by the Imperial Cancer Research Fund.

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vassaux, G., Hurst, H. & Lemoine, N. Insulation of a conditionally expressed transgene in an adenoviral vector. Gene Ther 6, 1192–1197 (1999). https://doi.org/10.1038/sj.gt.3300910

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3300910

Keywords

This article is cited by

Search

Quick links