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Increased persistence of lung gene expression using plasmids containing the ubiquitin C or elongation factor1α promoter

Gene Therapy volume 8, pages 1539–1546 (2001)Cite this article

Abstract

For effective gene therapy of chronic disease, persistent transgene expression at therapeutic levels is required. Clinical studies of airway gene transfer in patients with cystic fibrosis (CF) have resulted in short-lived transgene expression. We used intra-nasal dosing of naked plasmid DNA to the murine lung as a model for investigating the duration of airway gene transfer from a series of reporter expression plasmids. Transgene expression was transient when mediated by the viral promoters CMV, RSV and SV40, falling to less than 10% of peak expression after 2 weeks, although the presence of the adenoviral E4ORF3 gene in cis, resulted in extended duration of reporter activity from the CMV promoter. Transient expression from these promoters was not due to loss of the vector as determined by quantitative TaqMan PCR analysis. However, use of the promoters from the human polybiquitin C (UbC) and the elongation factor 1α (EF1α) genes resulted in persistent gene expression in the mouse lung. The UbC promoter directed high-level reporter activity which was maintained for up to 8 weeks and was still detectable 6 months after a single administration. Such persistent airway transgene expression from a nonviral vector without the concomitant expression of a potential antigen has not been reported previously. Thus, despite the persistence of vector DNA in vivo, attenuation of promoter function may lead to silencing of transgene expression and careful selection of promoter sequences is recommended for in vivo gene transfer.

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Acknowledgements

The authors thank Gary Ketner for the kind gift of anti-E4 ORF3 antibody and Brian Seed for plasmid πH3-CD8. The study was funded by the Cystic Fibrosis Trust (UK), Medical Research Council (UK), the Wellcome Trust and the National Lottery Fund.

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Authors and Affiliations

  1. Nuffield Department of Clinical Laboratory Sciences, GeneMedicine Research Group, University of Oxford, John Radcliffe Hospital, Oxford, OX3 9DU, UK

    DR Gill, SE Smyth & SC Hyde

  2. Department of Physiology, University of Cambridge, Downing Street, Cambridge, UK

    CA Goddard & WH Colledge

  3. MRC Clinical Sciences Centre, Imperial College School of Medicine, Hammersmith Hospital, London, UK

    CF Higgins

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  1. DR Gill
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Gill, D., Smyth, S., Goddard, C. et al. Increased persistence of lung gene expression using plasmids containing the ubiquitin C or elongation factor1α promoter. Gene Ther 8, 1539–1546 (2001). https://doi.org/10.1038/sj.gt.3301561

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