Molecular characterization, by digital PCR analysis of four HMBS gene mutations affecting the ubiquitous isoform of Porphobilinogen Deaminase (PBGD) in patients with Acute Intermittent Porphyria (AIP)

Francesca Granata, Manuel Mendez, Valentina Brancaleoni, Francisco J. Castelbon, Giovanna Graziadei, Paolo Ventura, Elena Di Pierro

Research output: Contribution to journalArticle

Abstract

Genetic variants in promoters and alternative-splicing lesions require to be experimentally tested in order to validate them as causatives of a disease. The digital PCR (dPCR) approach, which is an alternative to the classical qPCR, is an innovative and a more sensitive method for the detection and quantification of nucleic acids. In the present study, we identified four HMBS gene mutations affecting the ubiquitous isoform of porphobilinogen deaminase (PBGD) and established a dPCR protocol which would be able to detect the different transcripts of this gene. With the application of this method, we were able to characterize the functional roles of these four genetic variants, demonstrating that all these mutations were causatives of the non-erythroid variant of the acute intermittent porphyria (AIP) disease.

Original languageEnglish
Pages (from-to)295-301
JournalMolecular Genetics and Metabolism
Volume125
Issue number3
DOIs
Publication statusPublished - 2018

Fingerprint

Hydroxymethylbilane Synthase
Acute Intermittent Porphyria
Protein Isoforms
Genes
Polymerase Chain Reaction
Mutation
Alternative Splicing
Nucleic Acids

Keywords

  • Digital PCR
  • Gene expression
  • HMBS
  • Porphyria
  • Promoter variants
  • Splicing isoform

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Endocrinology

Cite this

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title = "Molecular characterization, by digital PCR analysis of four HMBS gene mutations affecting the ubiquitous isoform of Porphobilinogen Deaminase (PBGD) in patients with Acute Intermittent Porphyria (AIP)",
abstract = "Genetic variants in promoters and alternative-splicing lesions require to be experimentally tested in order to validate them as causatives of a disease. The digital PCR (dPCR) approach, which is an alternative to the classical qPCR, is an innovative and a more sensitive method for the detection and quantification of nucleic acids. In the present study, we identified four HMBS gene mutations affecting the ubiquitous isoform of porphobilinogen deaminase (PBGD) and established a dPCR protocol which would be able to detect the different transcripts of this gene. With the application of this method, we were able to characterize the functional roles of these four genetic variants, demonstrating that all these mutations were causatives of the non-erythroid variant of the acute intermittent porphyria (AIP) disease.",
keywords = "Digital PCR, Gene expression, HMBS, Porphyria, Promoter variants, Splicing isoform",
author = "Francesca Granata and Manuel Mendez and Valentina Brancaleoni and Castelbon, {Francisco J.} and Giovanna Graziadei and Paolo Ventura and {Di Pierro}, Elena",
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AU - Granata, Francesca

AU - Mendez, Manuel

AU - Brancaleoni, Valentina

AU - Castelbon, Francisco J.

AU - Graziadei, Giovanna

AU - Ventura, Paolo

AU - Di Pierro, Elena

PY - 2018

Y1 - 2018

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AB - Genetic variants in promoters and alternative-splicing lesions require to be experimentally tested in order to validate them as causatives of a disease. The digital PCR (dPCR) approach, which is an alternative to the classical qPCR, is an innovative and a more sensitive method for the detection and quantification of nucleic acids. In the present study, we identified four HMBS gene mutations affecting the ubiquitous isoform of porphobilinogen deaminase (PBGD) and established a dPCR protocol which would be able to detect the different transcripts of this gene. With the application of this method, we were able to characterize the functional roles of these four genetic variants, demonstrating that all these mutations were causatives of the non-erythroid variant of the acute intermittent porphyria (AIP) disease.

KW - Digital PCR

KW - Gene expression

KW - HMBS

KW - Porphyria

KW - Promoter variants

KW - Splicing isoform

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