A novel mutation deep within intron 7 of the GBA gene causes Gaucher disease
Abstract Background Mutations in the GBA gene that encodes the lysosomal enzyme acid β‐glucocerebrosidase cause Gaucher disease (GD), the most common lysosomal storage disorder. Most of the mutations are missense/nonsense, however, a few splicing mutations within or close to conserved consensus dono...
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doaj-ed28cb0f8c164a91bcb9899c190800d32020-11-25T00:28:46ZengWileyMolecular Genetics & Genomic Medicine2324-92692020-03-0183n/an/a10.1002/mgg3.1090A novel mutation deep within intron 7 of the GBA gene causes Gaucher diseaseAnna Malekkou0Ioanna Sevastou1Gavriella Mavrikiou2Theodoros Georgiou3Lluisa Vilageliu4Marina Moraitou5Helen Michelakakis6Chrystalla Prokopiou7Anthi Drousiotou8Department of Biochemical Genetics The Cyprus Institute of Neurology and Genetics Nicosia CyprusDepartment of Biochemical Genetics The Cyprus Institute of Neurology and Genetics Nicosia CyprusDepartment of Biochemical Genetics The Cyprus Institute of Neurology and Genetics Nicosia CyprusDepartment of Biochemical Genetics The Cyprus Institute of Neurology and Genetics Nicosia CyprusDepartment of Genetics Faculty of Biology Universitat de Barcelona IBUB, CIBERER Barcelona SpainDepartment of Enzymology and Cellular Function Institute of Child Health Athens GreeceDepartment of Enzymology and Cellular Function Institute of Child Health Athens GreeceDepartment of Haematology Limassol General Hospital Limassol CyprusDepartment of Biochemical Genetics The Cyprus Institute of Neurology and Genetics Nicosia CyprusAbstract Background Mutations in the GBA gene that encodes the lysosomal enzyme acid β‐glucocerebrosidase cause Gaucher disease (GD), the most common lysosomal storage disorder. Most of the mutations are missense/nonsense, however, a few splicing mutations within or close to conserved consensus donor or acceptor splice sites have also been described. The aim of the study was to identify the mutation(s) in a Cypriot patient with type I GD. Methods The genomic DNA of the proband was screened for nine common mutations using Polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) analysis. All exons and exon‐intron boundaries, and the 5’UTR and 3’UTR regions of the GBA gene, were investigated by Sanger sequencing. RNA analysis was performed using standard procedures, and the abnormal transcript was further cloned into pGEM‐T‐Easy plasmid vector and sequenced. The relevant intronic region was further sequenced by the Sanger method to identify the genetic variant. Results A novel point mutation, g.12599C > A (c.999 + 242C > A), was detected deep in intron 7 of the GBA gene. This type of mutation has been previously described for other diseases but this is the first time, as far as we know, that it is described for GD. This mutation creates a new donor splice site leading to aberrant splicing and resulting in the insertion of the first 239nt of intron 7 as a pseudoexon in the mRNA, creating a premature stop codon. Conclusion This study expands the mutation spectrum of GD and highlights the importance of RNA sequencing for the molecular diagnosis of patients bearing mutations in nonexonic regions.https://doi.org/10.1002/mgg3.1090Cypriotdeep intronic mutationGaucher diseaseGBAglucocerebrosidase |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Anna Malekkou Ioanna Sevastou Gavriella Mavrikiou Theodoros Georgiou Lluisa Vilageliu Marina Moraitou Helen Michelakakis Chrystalla Prokopiou Anthi Drousiotou |
spellingShingle |
Anna Malekkou Ioanna Sevastou Gavriella Mavrikiou Theodoros Georgiou Lluisa Vilageliu Marina Moraitou Helen Michelakakis Chrystalla Prokopiou Anthi Drousiotou A novel mutation deep within intron 7 of the GBA gene causes Gaucher disease Molecular Genetics & Genomic Medicine Cypriot deep intronic mutation Gaucher disease GBA glucocerebrosidase |
author_facet |
Anna Malekkou Ioanna Sevastou Gavriella Mavrikiou Theodoros Georgiou Lluisa Vilageliu Marina Moraitou Helen Michelakakis Chrystalla Prokopiou Anthi Drousiotou |
author_sort |
Anna Malekkou |
title |
A novel mutation deep within intron 7 of the GBA gene causes Gaucher disease |
title_short |
A novel mutation deep within intron 7 of the GBA gene causes Gaucher disease |
title_full |
A novel mutation deep within intron 7 of the GBA gene causes Gaucher disease |
title_fullStr |
A novel mutation deep within intron 7 of the GBA gene causes Gaucher disease |
title_full_unstemmed |
A novel mutation deep within intron 7 of the GBA gene causes Gaucher disease |
title_sort |
novel mutation deep within intron 7 of the gba gene causes gaucher disease |
publisher |
Wiley |
series |
Molecular Genetics & Genomic Medicine |
issn |
2324-9269 |
publishDate |
2020-03-01 |
description |
Abstract Background Mutations in the GBA gene that encodes the lysosomal enzyme acid β‐glucocerebrosidase cause Gaucher disease (GD), the most common lysosomal storage disorder. Most of the mutations are missense/nonsense, however, a few splicing mutations within or close to conserved consensus donor or acceptor splice sites have also been described. The aim of the study was to identify the mutation(s) in a Cypriot patient with type I GD. Methods The genomic DNA of the proband was screened for nine common mutations using Polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) analysis. All exons and exon‐intron boundaries, and the 5’UTR and 3’UTR regions of the GBA gene, were investigated by Sanger sequencing. RNA analysis was performed using standard procedures, and the abnormal transcript was further cloned into pGEM‐T‐Easy plasmid vector and sequenced. The relevant intronic region was further sequenced by the Sanger method to identify the genetic variant. Results A novel point mutation, g.12599C > A (c.999 + 242C > A), was detected deep in intron 7 of the GBA gene. This type of mutation has been previously described for other diseases but this is the first time, as far as we know, that it is described for GD. This mutation creates a new donor splice site leading to aberrant splicing and resulting in the insertion of the first 239nt of intron 7 as a pseudoexon in the mRNA, creating a premature stop codon. Conclusion This study expands the mutation spectrum of GD and highlights the importance of RNA sequencing for the molecular diagnosis of patients bearing mutations in nonexonic regions. |
topic |
Cypriot deep intronic mutation Gaucher disease GBA glucocerebrosidase |
url |
https://doi.org/10.1002/mgg3.1090 |
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