PUBLICATION
Generation and Characterization of a Zebrafish Model for ADGRV1-Associated Retinal Dysfunction Using CRISPR/Cas9 Genome Editing Technology
- Authors
- Stemerdink, M., Broekman, S., Peters, T., Kremer, H., de Vrieze, E., van Wijk, E.
- ID
- ZDB-PUB-230628-53
- Date
- 2023
- Source
- Cells 12(12): (Journal)
- Registered Authors
- de Vrieze, Erik, Kremer, Hannie, van Wijk, Erwin
- Keywords
- ADGRV1, CRISPR/Cas9, Usher syndrome, retinal dysfunction, retinitis pigmentosa, zebrafish
- MeSH Terms
-
- Animals
- CRISPR-Cas Systems/genetics
- Gene Editing
- Humans
- Retina
- Retinitis Pigmentosa*/genetics
- Zebrafish*/genetics
- PubMed
- 37371069 Full text @ Cells
Citation
Stemerdink, M., Broekman, S., Peters, T., Kremer, H., de Vrieze, E., van Wijk, E. (2023) Generation and Characterization of a Zebrafish Model for ADGRV1-Associated Retinal Dysfunction Using CRISPR/Cas9 Genome Editing Technology. Cells. 12(12):.
Abstract
Worldwide, around 40,000 people progressively lose their eyesight as a consequence of retinitis pigmentosa (RP) caused by pathogenic variants in the ADGRV1 gene, for which currently no treatment options exist. A model organism that mimics the human phenotype is essential to unravel the exact pathophysiological mechanism underlying ADGRV1-associated RP, and to evaluate future therapeutic strategies. The introduction of CRISPR/Cas-based genome editing technologies significantly improved the possibilities of generating mutant models in a time- and cost-effective manner. Zebrafish have been recognized as a suitable model to study Usher syndrome-associated retinal dysfunction. Using CRISPR/Cas9 technology we introduced a 4bp deletion in adgrv1 exon 9 (adgrv1rmc22). Immunohistochemical analysis showed that Adgrv1 was absent from the region of the photoreceptor connecting cilium in the adgrv1rmc22 zebrafish retina. Here, the absence of Adgrv1 also resulted in reduced levels of the USH2 complex members usherin and Whrnb, suggesting that Adgrv1 interacts with usherin and Whrnb in zebrafish photoreceptors. When comparing adgrv1rmc22 zebrafish with wild-type controls, we furthermore observed increased levels of aberrantly localized rhodopsin in the photoreceptor cell body, and decreased electroretinogram (ERG) B-wave amplitudes which indicate that the absence of Adgrv1 results in impaired retinal function. Based on these findings we present the adgrv1rmc22 zebrafish as the first ADGRV1 mutant model that displays an early retinal dysfunction. Moreover, the observed phenotypic changes can be used as quantifiable outcome measures when evaluating the efficacy of future novel therapeutic strategies for ADGRV1-associated RP.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping