The intronic ABCA4 c.5461-10T>C variant, frequently seen in patients with Stargardt disease, causes splice defects and reduced ABCA4 protein level.
Acta Ophthalmol
; 95(3): 240-246, 2017 May.
Article
de En
| MEDLINE
| ID: mdl-27775217
ABSTRACT
PURPOSE:
Despite being the third most common ABCA4 variant observed in patients with Stargardt disease, the functional effect of the intronic ABCA4 variant c.5461-10T>C is unknown. The purpose of this study was to investigate the molecular effect of this variant.METHODS:
Fibroblast samples from patients carrying the ABCA4 variant c.5461-10T>C were analysed by isolating total RNA, followed by real-time polymerase chain reaction (RT-PCR) using specific primers spanning the variant. For detection of ABCA4 protein, fibroblast samples were lysed and analysed by SDS-PAGE followed by immunoblotting using a monoclonal ABCA4 antibody.RESULTS:
The ABCA4 variant c.5461-10T>C causes a splicing defect resulting in the reduction of full-length mRNA in fibroblasts from patients and the presence of alternatively spliced mRNAs where exon 39-40 is skipped. A reduced level of full-length ABCA4 protein is observed compared to controls not carrying the variant.CONCLUSIONS:
This study describes the functional effect and the molecular mechanism of the pathogenic ABCA4 variant c.5461-10T>C. The variant is functionally important as it leads to splicing defects and a reduced level of ABCA4 protein.Mots clés
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
ARN
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Transporteurs ABC
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Dégénérescence maculaire
/
Mutation
Type d'étude:
Diagnostic_studies
/
Etiology_studies
Limites:
Adult
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Female
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Humans
/
Male
Langue:
En
Journal:
Acta Ophthalmol
Sujet du journal:
OFTALMOLOGIA
Année:
2017
Type de document:
Article
Pays d'affiliation:
Norvège