Loss-of-function mutations inUSH2Aare among the most common causes of syndromic and non-syndromic retinitis pigmentosa (RP). We previously presented skipping ofUSH2Aexon 13 as a promising treatment paradigm forUSH2A-associated RP. However, RP-associated mutations are often private, and evenly distributed along theUSH2Agene. In order to broaden the group of patients that could benefit from therapeutic exon skipping strategies, we expanded our approach to otherUSH2Aexons in which unique loss-of-function mutations have been reported by implementing a protein domain-oriented dual exon skipping strategy. We first generated zebrafish mutants carrying a genomic deletion of the orthologous exons of the frequently mutated humanUSH2Aexons 30–31 or 39–40 using CRISPR-Cas9. Excision of these in-frame combinations of exons restored usherin expression in the zebrafish retina and rescued the photopigment mislocalization typically observed inush2amutants. To translate these findings into a future treatment in humans, we employedin vitroassays to identify and validate antisense oligonucleotides (ASOs) with a high potency for sequence-specific dual exon skipping. Together, thein vitroandin vivodata demonstrate protein domain-oriented ASO-induced dual exon skipping to be a highly promising treatment option for RP caused by mutations inUSH2A.