The intronic ABCA4 c.5461-10TC variant, frequently seen in patients with Stargardt disease, causes splice defects and reduced ABCA4 protein level
| Autor: | Per M. Knappskog, Siren Berland, Cecilie Bredrup, Hilde E. Rusaas, Ingvild Aukrust, Gunnar Houge, Marte G. Haug, Agnete Jørgensen, Ragnhild Wivestad Jansson, Eyvind Rødahl |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Adult Male DNA Mutational Analysis Immunoblotting ABCA4 Retinal Pigment Epithelium Real-Time Polymerase Chain Reaction law.invention 03 medical and health sciences Exon Macular Degeneration Young Adult 0302 clinical medicine law medicine Electroretinography Humans Stargardt Disease splice Fibroblast Polymerase chain reaction Cells Cultured Genetics Messenger RNA biology General Medicine Exons Fibroblasts medicine.disease Rod Cell Outer Segment Molecular biology Introns Pedigree Stargardt disease Ophthalmology 030104 developmental biology medicine.anatomical_structure Phenotype RNA splicing Mutation 030221 ophthalmology & optometry biology.protein RNA ATP-Binding Cassette Transporters Female Tomography Optical Coherence |
| Zdroj: | Acta ophthalmologica. 95(3) |
| ISSN: | 1755-3768 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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