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Development of S/MAR minicircles for enhanced and persistent transgene expression in the mouse liver

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Abstract

We have previously described the development of a scaffold/matrix attachment region (S/MAR) episomal vector system for in vivo application and demonstrated its utility to sustain transgene expression in the mouse liver for at least 6 months following a single administration. Subsequently, we observed that transgene expression is sustained for the lifetime of the animal. The level of expression, however, does drop appreciably over time. We hypothesised that by eliminating the bacterial components in our vectors, we could improve their performance since bacterial sequences have been shown to be responsible for the immunotoxicity of the vector and the silencing of its expression when applied in vivo. We describe here the development of a minimally sized S/MAR vector, which is devoid of extraneous bacterial sequences. This minicircle vector comprises an expression cassette and an S/MAR moiety, providing higher and more sustained transgene expression for several months in the absence of selection, both in vitro and in vivo. In contrast to the expression of our original S/MAR plasmid vector, the novel S/MAR minicircle vectors mediate increased transgene expression, which becomes sustained at about twice the levels observed immediately after administration. These promising results demonstrate the utility of minimally sized S/MAR vectors for persistent, atoxic gene expression.

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Acknowledgements

The work was supported by the Myrovlytis Trust.

Conflict of interest

The authors declare no conflict of interest.

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Authors and Affiliations

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

    Orestis Argyros, Suet Ping Wong, Constantinos Fedonidis, Marcello Niceta, Charles Coutelle & Richard P. Harbottle

  2. Cardiovascular Science, Faculty of Medicine, National Heart and Lung Institute, Imperial College London, London, UK

    Oleg Tolmachov

  3. University College London, Institute for Women’s Health, London, UK

    Simon N. Waddington

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

    Steven J. Howe

Authors
  1. Orestis Argyros
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  2. Suet Ping Wong
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  3. Constantinos Fedonidis
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  4. Oleg Tolmachov
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  5. Simon N. Waddington
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  6. Steven J. Howe
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  7. Marcello Niceta
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  8. Charles Coutelle
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  9. Richard P. Harbottle
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Corresponding author

Correspondence to Richard P. Harbottle.

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Argyros, O., Wong, S.P., Fedonidis, C. et al. Development of S/MAR minicircles for enhanced and persistent transgene expression in the mouse liver. J Mol Med 89, 515–529 (2011). https://doi.org/10.1007/s00109-010-0713-3

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  • DOI: https://doi.org/10.1007/s00109-010-0713-3

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