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Aberrant RNA splicing is the major pathogenic effect in a knockāin mouse model of the dominantly inherited c.1430A>G humanRPE65mutation
- Source :
- Human Mutation. 40:426-443
- Publication Year :
- 2019
- Publisher :
- Hindawi Limited, 2019.
-
Abstract
- Human RPE65 mutations cause a spectrum of retinal dystrophies that result in blindness. While RPE65 mutations have been almost invariably recessively inherited, a c.1430A>G (p.(D477G)) mutation has been reported to cause autosomal dominant retinitis pigmentosa (adRP). To study the pathogenesis of this human mutation, we have replicated the mutation in a knock-in (KI) mouse model using CRISPR/Cas9-mediated genome editing. Significantly, in contrast to human patients, heterozygous KI mice do not exhibit any phenotypes in visual function tests. When raised in regular vivarium conditions, homozygous KI mice display relatively undisturbed visual functions with minimal retinal structural changes. However, KI/KI mouse retinae are more sensitive to light exposure and exhibit signs of degenerative features when subjected to light stress. We find that instead of merely producing a missense mutant protein, the A>G nucleotide substitution greatly affects appropriate splicing of Rpe65 mRNA by generating an ectopic splice site in comparable context to the canonical one, thereby disrupting RPE65 protein expression. Similar splicing defects were also confirmed for the human RPE65 c.1430G mutant in an in vitro Exontrap assay. Our data demonstrate that a splicing defect is associated with c.1430G pathogenesis, and therefore provide insights in the therapeutic strategy for human patients.
- Subjects :
- cis-trans-Isomerases
Genotype
RNA Splicing
Mutant
Gene Expression
Mice, Transgenic
Context (language use)
Biology
medicine.disease_cause
Retina
Article
Mice
03 medical and health sciences
Mutant protein
Genetics
medicine
Animals
Humans
Missense mutation
Genetic Predisposition to Disease
Alleles
Genetic Association Studies
Genetics (clinical)
030304 developmental biology
0303 health sciences
Mutation
Gene Expression Profiling
030305 genetics & heredity
Phenotype
Molecular biology
eye diseases
Disease Models, Animal
RPE65
RNA splicing
RNA Splice Sites
sense organs
Biomarkers
Subjects
Details
- ISSN :
- 10981004 and 10597794
- Volume :
- 40
- Database :
- OpenAIRE
- Journal :
- Human Mutation
- Accession number :
- edsair.doi.dedup.....a6a3e618a93af7d8fde50f4aa57918cc