Upstream open reading frames (uORFs) are fascinating translation regulatory elements that generally inhibit translation of the primary ORF. There is a huge knowledge gap in uORF regulation in the healthy retina and inherited retinal disease (IRD), a major cause of inherited blindness for which gene therapy is entering the clinic. By evaluating 4000 IRD patients, we identified a novel uORF-introducing mutation in the RDH12 gene in multiple families. In addition, we discovered naturally occurring uORFs in PRPH2, a major disease gene for autosomal dominant IRD with haploinsufficiency. We hypothesize that 1) uORFs play a role in retina-specific translational regulation, and 2) uORFs could represent a novel therapeutic target for IRD. To investigate this, we first aim to scrutinize the uORF landscape in the human retina by employing a combined approach of Ribo-seq and N-terminomics, followed by in-depth functional validation of selected uORFs. Secondly, we will explore antisense oligonucleotide (ASO) treatment to block recognition of the RDH12 and PRPH2 uORFs in both reporter assays and wild-type cellular models, as well as in patient-derived retinal organoids. Overall, this innovative study will provide major scientific insights in uORF-mediated translational regulation in the human retina and represents a unique proof of concept of uORF ASOs as novel genetic therapy with a potential broad application.