Next generation sequencing of pooled samples reveals new SNRNP200 mutations associated with retinitis pigmentosa.

Détails

ID Serval
serval:BIB_13C2C1338D52
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Next generation sequencing of pooled samples reveals new SNRNP200 mutations associated with retinitis pigmentosa.
Périodique
Human Mutation
Auteur(s)
Benaglio P., McGee T.L., Capelli L.P., Harper S., Berson E.L., Rivolta C.
ISSN
1098-1004 (Electronic)
ISSN-L
1059-7794
Statut éditorial
Publié
Date de publication
2011
Peer-reviewed
Oui
Volume
32
Numéro
6
Pages
E2246-E2258
Langue
anglais
Résumé
The gene SNRNP200 is composed of 45 exons and encodes a protein essential for pre-mRNA splicing, the 200 kDa helicase hBrr2. Two mutations in SNRNP200 have recently been associated with autosomal dominant retinitis pigmentosa (adRP), a retinal degenerative disease, in two families from China. In this work we analyzed the entire 35-Kb SNRNP200 genomic region in a cohort of 96 unrelated North American patients with adRP. To complete this large-scale sequencing project, we performed ultra high-throughput sequencing of pooled, untagged PCR products. We then validated the detected DNA changes by Sanger sequencing of individual samples from this cohort and from an additional one of 95 patients. One of the two previously known mutations (p.S1087L) was identified in 3 patients, while 4 new missense changes (p.R681C, p.R681H, p.V683L, p.Y689C) affecting highly conserved codons were identified in 6 unrelated individuals, indicating that the prevalence of SNRNP200-associated adRP is relatively high. We also took advantage of this research to evaluate the pool-and-sequence method, especially with respect to the generation of false positive and negative results. We conclude that, although this strategy can be adopted for rapid discovery of new disease-associated variants, it still requires extensive validation to be used in routine DNA screenings. © 2011 Wiley-Liss, Inc.
Pubmed
Web of science
Open Access
Oui
Création de la notice
29/06/2011 12:17
Dernière modification de la notice
20/08/2019 12:42
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