PUBLICATION
Exploring exon excision as a therapeutic intervention strategy for the future treatment of ADGRV1-associated retinitis pigmentosa
- Authors
- Stemerdink, M., Malinar, L., Broekman, S., Peters, T., Ensink, I., Ivanchenko, M.V., Venselaar, H., Kremer, H., de Vrieze, E., van Wijk, E.
- ID
- ZDB-PUB-251002-18
- Date
- 2025
- Source
- Molecular therapy. Nucleic acids 36: 102702102702 (Journal)
- Registered Authors
- de Vrieze, Erik, Kremer, Hannie, van Wijk, Erwin
- Keywords
- ADGRV1, CRISPR-Cas9, MT: RNA/DNA Editing, Usher syndrome type 2c, multi-exon excision, retinitis pigmentosa, therapies and applications, zebrafish
- MeSH Terms
- none
- PubMed
- 41036464 Full text @ Mol Ther Nucleic Acids
Citation
Stemerdink, M., Malinar, L., Broekman, S., Peters, T., Ensink, I., Ivanchenko, M.V., Venselaar, H., Kremer, H., de Vrieze, E., van Wijk, E. (2025) Exploring exon excision as a therapeutic intervention strategy for the future treatment of ADGRV1-associated retinitis pigmentosa. Molecular therapy. Nucleic acids. 36:102702102702.
Abstract
Given the lack of treatment options for ADGRV1-associated retinitis pigmentosa (RP), we evaluated exon excision as a potential therapeutic strategy. To enable careful selection of target exons, in silico analyses of the ADGRV1 protein domain structure were performed using AlphaFold2. These revealed a more complex ADGRV1 domain architecture compared to previous 2D assessments, indicating the presence of a larger number of Calxβ domains as well as a previously unidentified cysteine-rich domain. Based on 2D and 3D protein modeling and the presence of loss-of-function mutations, we selected two targets for exon excision: exon 9 and exons 40-42. To assess functional consequences, we generated zebrafish mutants lacking these exons using CRISPR-Cas9. While exon 9 excision failed to result in Adgrv1 expression, excision of exons 40-42 preserved protein expression, enabling proper USH2 protein complex assembly, rhodopsin trafficking, and electroretinogram (ERG) responses comparable to wild type. Additionally, CRISPR-Cas9-mediated removal of ADGRV1 exons 40-42 in HEK293T cells demonstrated the feasibility of this approach in the context of the human genome. These findings suggest that a carefully designed exon excision strategy may offer a viable treatment for ADGRV1-associated RP. Furthermore, they highlight the importance of 3D protein modeling in predicting structural consequences of exon excision and optimizing therapeutic design.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping