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Functional analysis and in vitro correction of splicing FAH mutations causing tyrosinemia type I

Authors

  • R. Pérez-Carro,

    1. Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular Severo Ochoa, Universidad Autónoma, CIBERER, IdiPaz, Madrid, Spain
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    • Current address: Departamento de Genética, Fundación Jiménez Diaz (IIS-FJD), Madrid, Spain

  • R. Sánchez-Alcudia,

    1. Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular Severo Ochoa, Universidad Autónoma, CIBERER, IdiPaz, Madrid, Spain
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    • Current address: Departamento de Genética, Fundación Jiménez Diaz (IIS-FJD), Madrid, Spain

  • B. Pérez,

    1. Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular Severo Ochoa, Universidad Autónoma, CIBERER, IdiPaz, Madrid, Spain
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  • R. Navarrete,

    1. Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular Severo Ochoa, Universidad Autónoma, CIBERER, IdiPaz, Madrid, Spain
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  • C. Pérez-Cerdá,

    1. Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular Severo Ochoa, Universidad Autónoma, CIBERER, IdiPaz, Madrid, Spain
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  • M. Ugarte,

    1. Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular Severo Ochoa, Universidad Autónoma, CIBERER, IdiPaz, Madrid, Spain
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  • L.R. Desviat

    Corresponding author
    1. Centro de Diagnóstico de Enfermedades Moleculares, Centro de Biología Molecular Severo Ochoa, Universidad Autónoma, CIBERER, IdiPaz, Madrid, Spain
    • Corresponding author: Lourdes R. Desviat, Centro de Biología Molecular Severo Ochoa, UAM-CSIC, Universidad Autónoma de Madrid, 28049 Madrid, Spain.

      Tel: +34 91 1964566;

      fax: +34 91 1964420;

      e-mail: lruiz@cbm.uam.es

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  • The authors declare no conflict of interest

Abstract

Hereditary tyrosinemia type I (HT1) is a rare disease caused by a deficiency of fumarylacetoacetate hydrolase (FAH) in the tyrosine catabolic pathway, resulting mainly in hepatic alterations due to accumulation of the toxic metabolites fumarylacetoacetate, maleylacetoacetate and succinylacetone. We have characterized using minigenes four splicing mutations affecting exonic or intronic nucleotides of the FAH gene identified in two HT1 patients. Two of the mutations are novel, c.82-1G>A and c.913G>C and the other two have been previously associated with a splicing defect (c.836A>G and c.1062+5G>A). All mutations were confirmed to affect splicing in minigenes, resulting in exon skipping or activation of a cryptic splice site. We have analyzed the effect of different compounds known to modulate splicing (valproic acid, phenyl butyrate, M344, EIPA, and resveratrol) and the overexpression of splice factors of the SR protein family on the transcriptional profile of the mutant minigenes. For the c.836A>G mutation, a partial recovery of the correctly spliced transcript was observed. These results confirm the relevance of performing functional studies for mutations potentially affecting the splicing process and open the possibility of supplementary therapeutic approaches to diseases caused by splicing defects.

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