title: Detection of novel genetic variation in autosomal dominant retinitis pigmentosa.
abstract: We explored an approach to detect disease-causing sequence variants in 448 candidate genes from five index cases of autosomal dominant retinitis pigmentosa (adRP) by sequence DNA capture and next-generation DNA sequencing (NGS). Detection of sequence variants was carried out by sequence capture NimbleGen and NGS in a SOLiD platform. After filtering out variants previously reported in genomic databases, novel potential adRP-causing variants were validated by dideoxy capillary electrophoresis (Sanger) sequencing and co-segregation in the families. A total of 55 novel sequence variants in the coding or splicing regions of adRP candidate genes were detected, 49 of which were confirmed by Sanger sequencing. Segregation of these variants in the corresponding adRP families showed three variants present in all the RP-affected members of the family. A novel mutation, p.L270R in IMPDH1, was found to be disease causing in one family. In another family a variant, p.M96T in the NRL gene was detected; this variant was previously reported as probably causing adRP. However, the previously reported p.A76V mutation in NRL as a cause of RP was excluded by co-segregation in the family. We discuss the benefits and limitations of our approach in the context of mutation detection in adRP patients.
authors.affiliation: Molecular Genetics Unit.
authors.forename: E
authors.initials: E
authors.lastname: Borràs
authors.forename: M
authors.initials: M
authors.lastname: de Sousa Dias
authors.forename: I
authors.initials: I
authors.lastname: Hernan
authors.forename: B
authors.initials: B
authors.lastname: Pascual
authors.forename: B
authors.initials: B
authors.lastname: Mañé
authors.forename: M J
authors.initials: MJ
authors.lastname: Gamundi
authors.forename: B
authors.initials: B
authors.lastname: Delás
authors.forename: M
authors.initials: M
authors.lastname: Carballo
mesh_terms: D050976:Basic-Leucine Zipper Transcription Factors / Q000235:genetics*; D005136:Eye Proteins / Q000235:genetics*; D005260:Female; D005799:Genes, Dominant; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007168:IMP Dehydrogenase / Q000235:genetics*; D040582:Inheritance Patterns; D008297:Male; D009154:Mutation; D016366:Open Reading Frames; D010375:Pedigree; D012326:RNA Splicing; D012174:Retinitis Pigmentosa / Q000235:genetics*
chemical_list: D050976:Basic-Leucine Zipper Transcription Factors; D005136:Eye Proteins; C490037:NRL protein, human; D007168:IMP Dehydrogenase; C488211:IMPDH1 protein, human
keywords: LR-PCR; autosomal dominant retinitis pigmentosa; next-generation sequencing; sequence capture