Cost-effective molecular inversion probe-based ABCA4 sequencing reveals deep-intronic variants in Stargardt disease
| العنوان: | Cost-effective molecular inversion probe-based ABCA4 sequencing reveals deep-intronic variants in Stargardt disease |
|---|---|
| المؤلفون: | Sem Bakker, Sabine Defoort, Eline Manders, Maartje van de Vorst, Mubeen Khan, Muhammad Imran Khan, Christian Gilissen, Duaa Elmelik, Frans P.M. Cremers, Ronny Derks, Aurore Devos, Felix Grassmann, Bernhard H. F. Weber, Claire Marie Dhaenens, Heidi Stöhr, Isabelle Meunier, Stéphanie S. Cornelis, Kornelia Neveling, Bernard Puech, Heidi L. Schulz |
| المصدر: | Human Mutation, 40, 1749-1759 Human Mutation, 40, 10, pp. 1749-1759 |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | Sequence analysis, RNA Splicing, DNA Mutational Analysis, ABCA4, Biology, Molecular Inversion Probe, DNA sequencing, Sensory disorders Donders Center for Medical Neuroscience [Radboudumc 12], 03 medical and health sciences, Exon, All institutes and research themes of the Radboud University Medical Center, Germany, Genetics, medicine, Humans, Stargardt Disease, splice, Genetic Predisposition to Disease, Genetics (clinical), Alleles, Genetic Association Studies, 030304 developmental biology, 0303 health sciences, 030305 genetics & heredity, Other Research Radboud Institute for Health Sciences [Radboudumc 0], Computational Biology, High-Throughput Nucleotide Sequencing, Molecular Sequence Annotation, Metabolic Disorders Radboud Institute for Molecular Life Sciences [Radboudumc 6], Exons, medicine.disease, Introns, Pedigree, Stargardt disease, Molecular Probes, RNA splicing, Mutation, biology.protein, ATP-Binding Cassette Transporters |
| الوصف: | Purpose Stargardt disease (STGD1) is caused by biallelic mutations in ABCA4, but many patients are genetically unsolved due to insensitive mutation-scanning methods. We aimed to develop a cost-effective sequencing method for ABCA4 exons and regions carrying known causal deep-intronic variants. Methods Fifty exons and 12 regions containing 14 deep-intronic variants of ABCA4 were sequenced using double-tiled single molecule Molecular Inversion Probe (smMIP)-based next-generation sequencing. DNAs of 16 STGD1 cases carrying 29 ABCA4 alleles and of four healthy persons were sequenced using 483 smMIPs. Thereafter, DNAs of 411 STGD1 cases with one or no ABCA4 variant were sequenced. The effect of novel noncoding variants on splicing was analyzed using in vitro splice assays. Results Thirty-four ABCA4 variants previously identified in 16 STGD1 cases were reliably identified. In 155/411 probands (38%), two causal variants were identified. We identified 11 deep-intronic variants present in 62 alleles. Two known and two new noncanonical splice site variants showed splice defects, and one novel deep-intronic variant (c.4539+2065C>G) resulted in a 170-nt mRNA pseudoexon insertion (p.[Arg1514Lysfs*35,=]). Conclusions smMIPs-based sequence analysis of coding and selected noncoding regions of ABCA4 enabled cost-effective mutation detection in STGD1 cases in previously unsolved cases. |
| تدمد: | 1059-7794 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::8b58b405c3dfa3546c7ed1fd19cbdc73 http://hdl.handle.net/2066/208474 |
| حقوق: | RESTRICTED |
| رقم الأكسشن: | edsair.doi.dedup.....8b58b405c3dfa3546c7ed1fd19cbdc73 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10597794 |
|---|