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Optimisation of electrotransfer of plasmid into skeletal muscle by pretreatment with hyaluronidase – increased expression with reduced muscle damage

Gene Therapy volume 8, pages 1264–1270 (2001)Cite this article

Abstract

The efficiency of plasmid gene transfer to skeletal muscle can be significantly improved by the application of an electrical field to the muscle following injection of plasmid DNA. However, this electrotransfer is associated with significant muscle damage which may result in substantial loss of transfected muscle fibres. Reduction of the voltage used in the technique can result in a decrease in muscle damage, with a concomitant reduction in expression, but without a significant decrease in the number of transfected fibres. Pre-treatment of the muscle with a solution of bovine hyaluronidase greatly increases the efficiency of plasmid gene transfer when used in conjunction with electrotransfer, but not when used alone. This combination treatment results in greatly enhanced levels of transfected muscle fibres without the increases in muscle damage associated with the electrotransfer process.

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Acknowledgements

We wish to thank George Dickson (Royal Holloway, London) and Jim Owen (Royal Free Hospital, London) for the loan of the BTX electroporator and electrodes. This work was funded by the Wellcome Trust and the Muscular Dystrophy Campaign. E Signori was supported by a short-term mobility fellowship from the Consiglio Nazionale delle Ricerche, Italy.

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Author notes

  1. JM McMahon and E Signori: The two first authors contributed equally to this work

Authors and Affiliations

  1. Department of Neuromuscular Diseases, Division of Neuroscience and Psychological Medicine, Gene Targeting Unit, Imperial College School of Medicine, Charing Cross Hospital, London, UK

    JM McMahon, KE Wells & DJ Wells

  2. Institute of Neurobiology and Molecular Medicine, CNR, Rome, Italy

    E Signori

  3. Laboratory of Molecular Pathology and Gene Therapy, IRCCS H ‘Casa Sollievo della Sofferenza’, San Giovanni Rotondo (FG), Rome, Italy

    E Signori & VM Fazio

  4. Laboratory for Molecular Medicine and Biotechnology, University Campus Bio-Medico, School of Medicine, Rome, Italy

    VM Fazio

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  1. JM McMahon
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  2. E Signori
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  3. KE Wells
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  4. VM Fazio
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  5. DJ Wells
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McMahon, J., Signori, E., Wells, K. et al. Optimisation of electrotransfer of plasmid into skeletal muscle by pretreatment with hyaluronidase – increased expression with reduced muscle damage. Gene Ther 8, 1264–1270 (2001). https://doi.org/10.1038/sj.gt.3301522

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  • DOI: https://doi.org/10.1038/sj.gt.3301522

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