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
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
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
Similar content being viewed by others
References
Douglas JT et al. Targeted gene delivery by tropism-modified adenoviral vectors Nature Biotech 1996 14: 1574–1578
Gutierrez AA, Lemoine NR, Sikora K . Gene therapy for cancer Lancet 1992 339: 715–721
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
Siders WM, Halloran PJ, Fenton RG . Transcriptional targeting of recombinant adenoviruses to human and murine melanoma cells Cancer Res 1996 56: 5638–5646
Rothmann T et al. Heart muscle-specific gene expression using replication defective recombinant adenovirus Gene Therapy 1996 3: 919–926
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
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
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
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
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
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
Hall PA et al. The c-erbB-2 proto-oncogene in human pancreatic cancer J Pathol 1990 161: 195–200
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
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
Martin LA, Lemoine NR . Direct cell killing by suicides genes Cancer Metas Rev 1996 15: 301–316
Mittereder N, March KL, Trapnell BC . Evaluation of the concentration and bioactivity of adenovirus vector for gene therapy J Virol 1996 70: 7498–7509
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
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
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
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
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
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
Rights 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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3300910
Keywords
This article is cited by
-
Enhanced antitumor efficacy of a novel oncolytic adenovirus combined with temozolomide in the treatment of melanoma in vivo
Journal of Cancer Research and Clinical Oncology (2015)
-
Targeting Gene-Viro-Therapy with AFP driving Apoptin gene shows potent antitumor effect in hepatocarcinoma
Journal of Biomedical Science (2012)
-
Modifications to the INSM1 promoter to preserve specificity and activity for use in adenoviral gene therapy of neuroendocrine carcinomas
Cancer Gene Therapy (2012)
-
Adenovirus-mediated hypoxia-targeted gene therapy using HSV thymidine kinase and bacterial nitroreductase prodrug-activating genes in vitro and in vivo
Cancer Gene Therapy (2011)
-
Cellular genetic tools to control oncolytic adenoviruses for virotherapy of cancer
Journal of Molecular Medicine (2008)