Zebrafish as a Model to Evaluate a CRISPR/Cas9-Based Exon Excision Approach as a Future Treatment Option for EYS-Associated Retinitis Pigmentosa

Autor: Theo A. Peters, Erik de Vrieze, Pam Graave, Sanne Broekman, Renske Schellens, Erwin van Wijk, Kerstin Nagel-Wolfrum, Rob W.J. Collin, Hannie Kremer
Rok vydání: 2021
Předmět:
QH301-705.5
Catalysis
Sensory disorders Donders Center for Medical Neuroscience [Radboudumc 12]
Article
Inorganic Chemistry
Exon
All institutes and research themes of the Radboud University Medical Center
EYS
Protein Domains
retinitis pigmentosa
Retinitis pigmentosa
medicine
CRISPR
Coding region
Animals
Biology (General)
Physical and Theoretical Chemistry
Outer nuclear layer
Eye Proteins
QD1-999
Molecular Biology
Zebrafish
CRISPR/Cas9
Spectroscopy
Genetics
exon skipping therapy
biology
Organic Chemistry
photoreceptors
General Medicine
Exons
Genetic Therapy
Zebrafish Proteins
medicine.disease
biology.organism_classification
zebrafish
Exon skipping
Computer Science Applications
Chemistry
Open reading frame
Disease Models
Animal
medicine.anatomical_structure
Phenotype
CRISPR-Cas Systems
antisense oligonucleotides
Zdroj: International Journal of Molecular Sciences
Volume 22
Issue 17
International Journal of Molecular Sciences, 22
International Journal of Molecular Sciences, 22, 17
International Journal of Molecular Sciences, Vol 22, Iss 9154, p 9154 (2021)
ISSN: 1422-0067
Popis: Retinitis pigmentosa (RP) is an inherited retinal disease (IRD) with an overall prevalence of 1 in 4000 individuals. Mutations in EYS (Eyes shut homolog) are among the most frequent causes of non-syndromic autosomal recessively inherited RP and act via a loss-of-function mechanism. In light of the recent successes for other IRDs, we investigated the therapeutic potential of exon skipping for EYS-associated RP. CRISPR/Cas9 was employed to generate zebrafish from which the region encompassing the orthologous exons 37-41 of human EYS (eys exons 40-44) was excised from the genome. The excision of these exons was predicted to maintain the open reading frame and to result in the removal of exactly one Laminin G and two EGF domains. Although the eysΔexon40-44 transcript was found at levels comparable to wild-type eys, and no unwanted off-target modifications were identified within the eys coding sequence after single-molecule sequencing, EysΔexon40-44 protein expression could not be detected. Visual motor response experiments revealed that eysΔexon40-44 larvae were visually impaired and histological analysis revealed a progressive degeneration of the retinal outer nuclear layer in these zebrafish. Altogether, the data obtained in our zebrafish model currently provide no indications for the skipping of EYS exons 37-41 as an effective future treatment strategy for EYS-associated RP.
Databáze: OpenAIRE
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