Abstract
Stargardt disease (STGD1), due to mutations in the large ABCA4 gene, is the most common inherited macular degeneration in humans. Attempts at developing gene therapy approaches for treatment of STGD1 are currently ongoing. Among all the vectors available for gene therapy of inherited retinal diseases, those based on adeno-associated viruses (AAV) are the most promising given the efficacy shown in various animal models and their excellent safety profile in humans, as confirmed in many ongoing clinical trials. However, one of the main obstacles for the use of AAV is their limited effective packaging capacity of about 5 kb. Taking advantage of the AAV genome’s ability to concatemerize , others and we have recently developed dual AAV vectors to overcome this limit. We tested dual AAV vectors for ABCA4 delivery, and found that they transduce efficiently both mouse and pig photoreceptors , and rescue the Abca4−/− mouse retinal phenotype, indicating their potential for gene therapy of STGD1. This chapter details how we designed dual AAV vectors for the delivery of the ABCA4 gene and describes the techniques that can be explored to evaluate dual AAV transduction efficiency in vitro and in the retina, and their efficacy in the mouse model of STGD1.
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Acknowledgments
Thanks to Raffaele Castello (Scientific Office, TIGEM, Pozzuoli, Italy) for the critical reading of this manuscript. Funding: The work was supported by the following: the European Research Council/ERC Grant agreement no282085 RetGeneTx; the European Community’s Seventh Framework Programme (FP7/2007–2013) under Grant agreement no 242013 Treatrush; the NIH (grant R24 EY019861-01A); the Italian Telethon Foundation (grant TGM11MT1).
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Trapani, I. (2018). Dual AAV Vectors for Stargardt Disease. In: Boon, C., Wijnholds, J. (eds) Retinal Gene Therapy. Methods in Molecular Biology, vol 1715. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7522-8_11
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DOI: https://doi.org/10.1007/978-1-4939-7522-8_11
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