Khan, Mubeen and Cornelis, Stephanie S. and Khan, Muhammad Imran and Elmelik, Duaa and Manders, Eline and Bakker, Sem and Derks, Ronny and Neveling, Kornelia and van DeVorst, Maartje and Gilissen, Christian and Meunier, Isabelle and Defoort, Sabine and Puech, Bernard and Devos, Aurore and Schulz, Heidi L. and Stoehr, Heidi and Grassmann, Felix and Weber, Bernhard H. F. and Dhaenens, Claire-Marie and Cremers, Frans P. M. (2019) Cost-effective molecular inversion probe-based ABCA4 sequencing reveals deep-intronic variants in Stargardt disease. HUMAN MUTATION, 40 (10). pp. 1749-1759. ISSN 1059-7794, 1098-1004
Full text not available from this repository. (Request a copy)Abstract
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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | CONE-ROD DYSTROPHY; SYNONYMOUS VARIANTS; RETINAL DISEASE; GENE; MUTATION; METAANALYSIS; ABCA4; deep-intronic variants; next generation sequencing; smMIPs; Stargardt disease |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Humangenetik |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 26 Mar 2020 13:45 |
| Last Modified: | 26 Mar 2020 13:45 |
| URI: | https://pred.uni-regensburg.de/id/eprint/26161 |
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