title: Antisense Oligonucleotide-based Splice Correction for USH2A-associated Retinal Degeneration Caused by a Frequent Deep-intronic Mutation.
abstract: Usher syndrome (USH) is the most common cause of combined deaf-blindness in man. The hearing loss can be partly compensated by providing patients with hearing aids or cochlear implants, but the loss of vision is currently untreatable. In general, mutations in the USH2A gene are the most frequent cause of USH explaining up to 50% of all patients worldwide. The first deep-intronic mutation in the USH2A gene (c.7595-2144A>G) was reported in 2012, leading to the insertion of a pseudoexon (PE40) into the mature USH2A transcript. When translated, this PE40-containing transcript is predicted to result in a truncated non-functional USH2A protein. In this study, we explored the potential of antisense oligonucleotides (AONs) to prevent aberrant splicing of USH2A pre-mRNA as a consequence of the c.7595-2144A>G mutation. Engineered 2'-O-methylphosphorothioate AONs targeting the PE40 splice acceptor site and/or exonic splice enhancer regions displayed significant splice correction potential in both patient derived fibroblasts and a minigene splice assay for USH2A c.7595-2144A>G, whereas a non-binding sense oligonucleotide had no effect on splicing. Altogether, AON-based splice correction could be a promising approach for the development of a future treatment for USH2A-associated retinitis pigmentosa caused by the deep-intronic c.7595-2144A>G mutation.
authors: Department of Otorhinolaryngology, Radboudumc, Nijmegen, the Netherlands.|Radboud Institute for Molecular Life Sciences, Radboudumc, Nijmegen, the Netherlands.
authors: Radulfus Wn
authors: RW
authors: Slijkerman
authors: Laboratoire de Génétique Moléculaire, CHU Montpellier, Montpellier, France.|Laboratoire de Génétique de Maladies Rares EA 7402, Université de Montpellier, Montpellier, France.
authors: Christel
authors: C
authors: Vaché
authors: Department of Otorhinolaryngology, Radboudumc, Nijmegen, the Netherlands.|Radboud Institute for Molecular Life Sciences, Radboudumc, Nijmegen, the Netherlands.
authors: Margo
authors: M
authors: Dona
authors: Laboratoire de Génétique de Maladies Rares EA 7402, Université de Montpellier, Montpellier, France.
authors: Gema
authors: G
authors: García-García
authors: Laboratoire de Génétique Moléculaire, CHU Montpellier, Montpellier, France.|Laboratoire de Génétique de Maladies Rares EA 7402, Université de Montpellier, Montpellier, France.
authors: Mireille
authors: M
authors: Claustres
authors: Department of Otorhinolaryngology, Radboudumc, Nijmegen, the Netherlands.|Donders Institute for Brain, Cognition, and Behaviour, Nijmegen, the Netherlands.
authors: Lisette
authors: L
authors: Hetterschijt
authors: Department of Otorhinolaryngology, Radboudumc, Nijmegen, the Netherlands.|Donders Institute for Brain, Cognition, and Behaviour, Nijmegen, the Netherlands.
authors: Theo A
authors: TA
authors: Peters
authors: Department of Otorhinolaryngology, Radboudumc, Nijmegen, the Netherlands.|Radboud Institute for Health Sciences, Radboudumc, Nijmegen, the Netherlands.
authors: Bas P
authors: BP
authors: Hartel
authors: Department of Otorhinolaryngology, Radboudumc, Nijmegen, the Netherlands.|Donders Institute for Brain, Cognition, and Behaviour, Nijmegen, the Netherlands.
authors: Ronald Je
authors: RJ
authors: Pennings
authors: Grupo de Investigación en Biomedicina Molecular, Celular y Genómica, and CIBERER, Madrid, Spain.
authors: José M
authors: JM
authors: Millan
authors: Grupo de Investigación en Biomedicina Molecular, Celular y Genómica, and CIBERER, Madrid, Spain.
authors: Elena
authors: E
authors: Aller
authors: Donders Institute for Brain, Cognition, and Behaviour, Nijmegen, the Netherlands.|Department of Human Genetics, Radboudumc, Nijmegen, the Netherlands.
authors: Alejandro
authors: A
authors: Garanto
authors: Donders Institute for Brain, Cognition, and Behaviour, Nijmegen, the Netherlands.|Department of Human Genetics, Radboudumc, Nijmegen, the Netherlands.
authors: Rob Wj
authors: RW
authors: Collin
authors: Department of Otorhinolaryngology, Radboudumc, Nijmegen, the Netherlands.|Donders Institute for Brain, Cognition, and Behaviour, Nijmegen, the Netherlands.|Department of Human Genetics, Radboudumc, Nijmegen, the Netherlands.
authors: Hannie
authors: H
authors: Kremer
authors: Laboratoire de Génétique Moléculaire, CHU Montpellier, Montpellier, France.|Laboratoire de Génétique de Maladies Rares EA 7402, Université de Montpellier, Montpellier, France.
authors: Anne-Françoise
authors: AF
authors: Roux
authors: Department of Otorhinolaryngology, Radboudumc, Nijmegen, the Netherlands.|Donders Institute for Brain, Cognition, and Behaviour, Nijmegen, the Netherlands.
authors: Erwin
authors: E
authors: Van Wijk