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Persistent episomal transgene expression in liver following delivery of a scaffold/matrix attachment region containing non-viral vector

Gene Therapy volume 15, pages 1593–1605 (2008)Cite this article

Abstract

An ideal gene therapy vector should enable persistent transgene expression without limitations of safety and reproducibility. Here we report the development of a non-viral episomal plasmid DNA (pDNA) vector that appears to fulfil these criteria. This pDNA vector combines a scaffold/matrix attachment region (S/MAR) with a human liver-specific promoter (α1-antitrypsin (AAT)) in such a way that long-term expression is enabled in murine liver following hydrodynamic injection. Long-term expression is demonstrated by monitoring the longitudinal luciferase expression profile for up to 6 months by means of in situ bioluminescent imaging. All relevant control pDNA constructs expressing luciferase are unable to sustain significant transgene expression beyond 1 week post-administration. We establish that this shutdown of expression is due to promoter methylation. In contrast, the S/MAR element appears to inhibit methylation of the AAT promoter thereby preventing transgene silencing. Although this vector appears to be maintained as an episome throughout, we have no evidence for its establishment as a replicating entity. We conclude that the combination of a mammalian, tissue-specific promoter with the S/MAR element is sufficient to drive long-term episomal pDNA expression of genes in vivo.

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Abbreviations

AAT:

α1-antitrypsin

CMV:

cytomegalovirus

pDNA:

plasmid DNA

S/MAR:

scaffold/matrix attachment region.

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Acknowledgements

The work was supported by the Medical Research Council. SNW is a Philip Gray Fellow of the Katharine Dormandy Trust.

Author information

Authors and Affiliations

  1. Department of Chemistry, Imperial College Genetic Therapies Centre, Imperial College London, London, UK

    O Argyros, A D Miller & R P Harbottle

  2. Gene Therapy Research Group, Section of Molecular Medicine, National Heart and Lung Institute, Imperial College London, London, UK

    S P Wong, M Niceta, C Coutelle & R P Harbottle

  3. Molecular Department of Haematology, Haemophilia Centre and Haemostasis Unit, Royal Free and University College Medical School, London, UK

    S N Waddington

  4. Molecular Immunology Unit, Institute of Child Health, University College London, London, UK

    S J Howe

  5. ImuThes Ltd, London, UK

    A D Miller

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  1. O Argyros
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Correspondence to A D Miller.

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Supplementary Information accompanies the paper on Gene Therapy website (http://www.nature.com/gt)

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Argyros, O., Wong, S., Niceta, M. et al. Persistent episomal transgene expression in liver following delivery of a scaffold/matrix attachment region containing non-viral vector. Gene Ther 15, 1593–1605 (2008). https://doi.org/10.1038/gt.2008.113

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