Functional analysis of synonymous substitutions predicted to affect splicing of the CFTR gene.
J Cyst Fibros
; 11(6): 511-7, 2012 Dec.
Article
em En
| MEDLINE
| ID: mdl-22591852
ABSTRACT
BACKGROUND:
Cystic fibrosis is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Over 1800 CFTR mutations have been reported, and about 12% of mutations are believed to impair pre-mRNA splicing. Given that several synthetic, non-splice-junction synonymous substitutions have been reported to alter splicing in CFTR, we predicted that naturally occurring synonymous substitutions may be erroneously classified as functionally neutral.METHODS:
Computational tools were used to predict the effect of synonymous substitutions on CFTR pre-mRNA splicing. The functional consequences of selected substitutions were evaluated using a minigene splicing assay.RESULTS:
Two synonymous mutations were shown to have a dramatic effect on CFTR pre-mRNA splicing, and consequently could alter protein integrity and phenotypic outcome.CONCLUSIONS:
Traditional methods of mutation analysis overlook splicing defects that occur at internal positions in coding exons, especially synonymous substitutions. We show that bioinformatics tools and minigene splicing assays are a potent combination to prioritize and identify mutations that cause aberrant CFTR pre-mRNA splicing.
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Análise Mutacional de DNA
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Processamento Alternativo
/
Regulador de Condutância Transmembrana em Fibrose Cística
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Sítios de Splice de RNA
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Fibrose Cística
Tipo de estudo:
Prognostic_studies
/
Risk_factors_studies
Limite:
Humans
Idioma:
En
Revista:
J Cyst Fibros
Ano de publicação:
2012
Tipo de documento:
Article
País de afiliação:
Estados Unidos