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AAV-mediated knockdown of Peripherin-2 in vivo using miRNA-based hairpins

Gene Therapy volume 17, pages 486–493 (2010)Cite this article

Abstract

Gene therapy for inherited retinal degeneration in which expression of a mutant allele has a gain-of-function effect on photoreceptor cells is likely to depend on efficient silencing of the mutated allele. Peripherin-2 (Prph2, also known as peripherin/RDS) is an abundantly expressed photoreceptor-specific gene. In humans, gain-of-function mutations in PRPH2 result in both autosomal dominant retinitis pigmentosa and dominant maculopathies. Gene-silencing strategies for these conditions include RNA interference by short hairpin RNAs (shRNAs). Recent evidence suggests that microRNA (miRNA)-based hairpins may offer a safer and more effective alternative. In this study, we used for the first time a virally transferred miRNA-based hairpin to silence Prph2 in the murine retina. The results show that an miRNA-based shRNA can efficiently and specifically silence Prph2 in vivo as early as 3 weeks after AAV2/8-mediated subretinal delivery, leading to a nearly 50% reduction of photoreceptor cells after 5 weeks. We conclude that miRNA-based hairpins can achieve rapid and robust gene silencing after efficient vector-mediated delivery to the retina. The rationale of using an miRNA-based template to improve the silencing efficiency of a hairpin may prove valuable for allele-specific silencing in which the choice for an RNAi target is limited and offers an alternative therapeutic strategy for the treatment of dominant retinopathies.

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Acknowledgements

We thank Dr Peter MG Munro for electron microscopy. This project was supported by the European Union (Integrated Project EVI-GENORET: LSHG-CT-2005-512036), Medical Research Council and the National Institute for Health Research Biomedical Centre for Ophthalmology. JWBB is a Wellcome Trust Advanced Fellow.

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Authors and Affiliations

  1. Department of Molecular Therapy, Institute of Ophthalmology, University College London, London, UK

    A Georgiadis, M Tschernutter, J W B Bainbridge, S J Robbie, A J Smith & R R Ali

  2. Department of Haematology, University College London, London, UK

    J McIntosh & A C Nathwani

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  1. A Georgiadis
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  2. M Tschernutter
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  3. J W B Bainbridge
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Correspondence to R R Ali.

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Georgiadis, A., Tschernutter, M., Bainbridge, J. et al. AAV-mediated knockdown of Peripherin-2 in vivo using miRNA-based hairpins. Gene Ther 17, 486–493 (2010). https://doi.org/10.1038/gt.2009.162

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