Study of USH1 Splicing Variants through Minigenes and Transcript Analysis from Nasal Epithelial Cells
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Study of USH1 Splicing Variants through Minigenes and Transcript Analysis from Nasal Epithelial Cells

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Study of USH1 Splicing Variants through Minigenes and Transcript Analysis from Nasal Epithelial Cells

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dc.contributor.author Aparisi Navarro, María José es_ES
dc.contributor.author García-García, Gema es_ES
dc.contributor.author Aller Mañas, Elena es_ES
dc.contributor.author Sequedo, María Dolores es_ES
dc.contributor.author Martínez-Fernández de la Cámara, Cristina es_ES
dc.contributor.author Rodrigo, Regina es_ES
dc.contributor.author Armengot Carceller, Miguel es_ES
dc.contributor.author Cortijo Gimeno, Julio es_ES
dc.contributor.author Milara Payá, Javier es_ES
dc.contributor.author Díaz Llopis, Manuel es_ES
dc.contributor.author Jaijo, Teresa es_ES
dc.contributor.author Millán Salvador, José María es_ES
dc.date.accessioned 2015-06-19T07:47:46Z
dc.date.available 2015-06-19T07:47:46Z
dc.date.issued 2013 es_ES
dc.identifier.uri http://hdl.handle.net/10550/44513
dc.description.abstract Usher syndrome type I (USH1) is an autosomal recessive disorder characterized by congenital profound deafness, vestibular areflexia and prepubertal retinitis pigmentosa. The first purpose of this study was to determine the pathologic nature of eighteen USH1 putative splicing variants found in our series and their effect in the splicing process by minigene assays. These variants were selected according to bioinformatic analysis. The second aim was to analyze the USH1 transcripts, obtained from nasal epithelial cells samples of our patients, in order to corroborate the observed effect of mutations by minigenes in patient’s tissues. The last objective was to evaluate the nasal ciliary beat frequency in patients with USH1 and compare it with control subjects. In silico analysis were performed using four bioinformatic programs: NNSplice, Human Splicing Finder, NetGene2 and Spliceview. Afterward, minigenes based on the pSPL3 vector were used to investigate the implication of selected changes in the mRNA processing. To observe the effect of mutations in the patient’s tissues, RNA was extracted from nasal epithelial cells and RT-PCR analyses were performed. Four MYO7A (c.470G>A, c.1342_1343delAG, c.5856G>A and c.3652G>A), three CDH23 (c.2289+1G>A, c.6049G>A and c.8722+1delG) and one PCDH15 (c.3717+2dupTT) variants were observed to affect the splicing process by minigene assays and/or transcripts analysis obtained from nasal cells. Based on our results, minigenes are a good approach to determine the implication of identified variants in the mRNA processing, and the analysis of RNA obtained from nasal epithelial cells is an alternative method to discriminate neutral Usher variants from those with a pathogenic effect on the splicing process. In addition, we could observe that the nasal ciliated epithelium of USH1 patients shows a lower ciliary beat frequency than control subjects. es_ES
dc.source PLoS ONE Vol. 8 Issue 2: es_ES
dc.title Study of USH1 Splicing Variants through Minigenes and Transcript Analysis from Nasal Epithelial Cells es_ES
dc.type info:eu-repo/semantics/article es_ES
dc.identifier.doi 10.1371/journal.pone.0057506 es_ES
dc.identifier.idgrec 102800 es_ES

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