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Erwin van Wijk, Margo Dona, Ralph Slijkerman, Peter Adamson, Janne Turunen, Maarten Kamermans, Stephan C F Neuhauss, Hester van Diepen; Antisense Oligonucleotide-induced Skipping of USH2A exon13 Restores Visual Function in Zebrafish. Invest. Ophthalmol. Vis. Sci. 2017;58(8):2490.
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Mutations in USH2A exon13 are the most frequent cause of both syndromic and non-syndromic retinitis pigmentosa (RP), for which currently no treatment options exist. It is generally believed that RP due to mutations in this gene is caused by a loss-of-function mechanism. Zebrafish lacking Ush2a show early signs of retinal dysfunction, although its regeneration capacity blocks the progression of retinal degeneration. Skipping of in-frame exons, like USH2A exon13, that carry loss-of-function mutations, will restore the open reading frame and potentially results in a slightly shortened protein with residual function. Therefore we explored the therapeutic potential of exon13-skipping as a therapeutic approach for future treatment of USH2A-associated retinal degeneration using zebrafish as a model.
Zebrafish larvae carrying a homozygous lesion (c.2337_2342delinsAC; p.Cys780Glnfs*) in Ush2a exon13, were injected with tailor-made antisense oligonucleotides (AONs) targeting this exon. Exon-skipping efficiency was determined by RT-PCR analysis and restoration of Ush2a protein expression and visual function was monitored by immunohistochemistry and electroretinogram (ERG) recordings, respectively.
Injection of morpholino-based AONs in homozygous mutant zebrafish larvae successfully induced skipping of exon13 from the mature ush2a mRNA. As a result, Ush2a protein expression at the photoreceptor periciliary membrane was partly restored. In addition, ERG traces were restored in AON-treated larvae as compared to uninjected mutant controls.
Proof-of-concept has been obtained for exon-skipping as a therapeutic approach for the development of a future treatment for USH2A-associated retinal degeneration caused by loss-of-function mutations in exon13.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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