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| Research Article |
| Optimisation of electrotransfer of plasmid into skeletal muscle by pretreatment with hyaluronidase - increased expression with reduced muscle damage |
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| J M McMahon1,a, E Signori2,3,a, K E Wells1, V M Fazio3,4 and D J Wells1 |
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1Gene Targeting Unit, Department of Neuromuscular Diseases, Division of Neuroscience and Psychological Medicine, Imperial College School of Medicine, Charing Cross Hospital, London, UK
2Institute of Neurobiology and Molecular Medicine, CNR, Rome, Italy
3Laboratory of Molecular Pathology and Gene Therapy, IRCCS H 'Casa Sollievo della Sofferenza', San Giovanni Rotondo (FG), Italy
4Laboratory for Molecular Medicine and Biotechnology, University Campus Bio-Medico, School of Medicine, Rome, Italy
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Correspondence to: D J Wells, Gene Targeting Unit, Department of Neuromuscular Diseases, Division of Neuroscience and Psychological Medicine, Imperial College School of Medicine, Charing Cross Hospital, St Dunstan's Road, London W6 8RP, UK
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aThe two first authors contributed equally to this work |
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| 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. Gene Therapy (2001) 8, 1264-1270. |
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| Keywords |
| muscular dystrophy; gene therapy; plasmid; skeletal muscle; electroporation; fibres |
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| Received 26 February 2001; accepted 11 June 2001 |
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| August 2001, Volume 8, Number 16, Pages 1264-1270 |
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