Repository logo
 
Publication

Unraveling the effect of silent, intronic and missense mutations on VWF splicing: contribution of next generation sequencing in the study of mRNA

2018-10-25Journal article Open access
Simple item page
dc.contributor.authorBorràs, N
dc.contributor.authorOrriols, G
dc.contributor.authorBatlle, J
dc.contributor.authorPérez-Rodríguez, A
dc.contributor.authorFidalgo, T
dc.contributor.authorMartinho, P
dc.contributor.authorLópez-Fernández, MF
dc.contributor.authorRodríguez-Trillo, Á
dc.contributor.authorLourés, E
dc.contributor.authorParra, R
dc.contributor.authorAltisent, C
dc.contributor.authorCid, AR
dc.contributor.authorBonanad, S
dc.contributor.authorCabrera, N
dc.contributor.authorMoret, A
dc.contributor.authorMingot-Castellano, ME
dc.contributor.authorNavarro, N
dc.contributor.authorPérez-Montes, R
dc.contributor.authorMarcellini, S
dc.contributor.authorMoreto, A
dc.contributor.authorHerrero, S
dc.contributor.authorSoto, I
dc.contributor.authorFernández-Mosteirín, N
dc.contributor.authorJiménez-Yuste, V
dc.contributor.authorAlonso, N
dc.contributor.authorde Andrés-Jacob, A
dc.contributor.authorFontanes, E
dc.contributor.authorCampos, R
dc.contributor.authorPaloma, MJ
dc.contributor.authorBermejo, N
dc.contributor.authorBerrueco, R
dc.contributor.authorMateo, J
dc.contributor.authorArribalzaga, K
dc.contributor.authorMarco, P
dc.contributor.authorPalomo, Á
dc.contributor.authorCastro Quismondo, N
dc.contributor.authorIñigo, B
dc.contributor.authorNieto, MM
dc.contributor.authorVidal, R
dc.contributor.authorMartínez, MP
dc.contributor.authorAguinaco, R
dc.contributor.authorTenorio, JM
dc.contributor.authorFerreiro, M
dc.contributor.authorGarcía-Frade, J
dc.contributor.authorRodríguez-Huerta, AM
dc.contributor.authorCuesta, J
dc.contributor.authorRodríguez-González, R
dc.contributor.authorGarcía-Candel, F
dc.contributor.authorDobón, M
dc.contributor.authorAguilar, C
dc.contributor.authorVidal, F
dc.contributor.authorCorrales, I
dc.date.accessioned2018-11-28T13:01:11Z
dc.date.available2018-11-28T13:01:11Z
dc.date.issued2018-10-25
dc.description.abstractLarge studies in von Willebrand disease patients, including Spanish and Portuguese registries, led to identification of >250 different mutations. It is a challenge to determine the pathogenic effect of potential splice site mutations on VWF mRNA. This study aimed to elucidate the true effects of 18 mutations on VWF mRNA processing, investigate the contribution of next-generation sequencing to in vivo mRNA study in von Willebrand disease, and compare the findings with in silico prediction. RNA extracted from patient platelets and leukocytes was amplified by RT-PCR and sequenced using Sanger and next generation sequencing techniques. Eight mutations affected VWF splicing: c.1533+1G>A, c.5664+2T>C and c.546G>A (p.=) prompted exon skipping; c.3223-7_3236dup and c.7082-2A>G resulted in activation of cryptic sites; c.3379+1G>A and c.7473G>A (p.=) demonstrated both molecular pathogenic mechanisms simultaneously; and the p.Cys370Tyr missense mutation generated two aberrant transcripts. Of note, the complete effect of 3 mutations was provided by next generation sequencing alone because of low expression of the aberrant transcripts. In the remaining 10 mutations, no effect was elucidated in the experiments. However, the differential findings obtained in platelets and leukocytes provided substantial evidence that 4 of these would have an effect on VWF levels. In this first report using next generation sequencing technology to unravel the effects of VWF mutations on splicing, the technique yielded valuable information. Our data bring to light the importance of studying the effect of synonymous and missense mutations on VWF splicing to improve the current knowledge of the molecular mechanisms behind von Willebrand disease.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationHaematologica. 2018 Oct 25. pii: haematol.2018.203166.pt_PT
dc.identifier.doi10.3324/haematol.2018.203166pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.4/2182
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.subjectMutaçãopt_PT
dc.subjectProcessamento de RNApt_PT
dc.titleUnraveling the effect of silent, intronic and missense mutations on VWF splicing: contribution of next generation sequencing in the study of mRNApt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.startPagehaematol.2018.203166pt_PT
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
Haematologica.pdf
Size:
4.3 MB
Format:
Adobe Portable Document Format
Download
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.71 KB
Format:
Item-specific license agreed upon to submission
Description:
Download

Collections

HEM- Artigos