Genetic analysis in cystic fibrosis (CF) is a difficult task. Within the many causes of variability and uncertainty, a major determinant is poor knowledge of the functional effect of most DNA variants of the Cystic Fibrosis Transmembrane conductance Regulator (CFTR) gene. In turn, knowledge of the effect of a CFTR variant has dramatic diagnostic, prognostic and, in the era of CF precision medicine, also therapeutic consequences. One of the most challenging CFTR variants is the (TG)mTn haplotype, which has variable functional effect and controversial clinical consequences. The exact quantification of the anomalous splicing of CFTR exon 10 (in the HGVS name; exon 9 in the legacy name) and, consequently, of the residual wild-type functional CFTR mRNA, should be mandatory in clinical assessment of patients with potentially pathological haplotype of this tract. Here, we present a real time-based assay for the quantification of the proportion of exon 10+/exon 10- CFTR mRNA, starting from nasal brushing. Our assay proved rapid, economic and easy to perform. Specific primers used for this assay are either disclosed or commercially available, allowing any laboratory to easily perform it. A simplified analysis of the data is provided, facilitating the interpretation of the results. This method helps to enhance the comprehension of the genotype- phenotype relationship in CF and CFTR-related disorders (CFTR-RD), crucial for the diagnosis, prognosis and personalized therapy of CF.

Quantitative evaluation of CFTR pre-mRNA splicing dependent on the (TG)mTn poly-variant tract / Sterrantino, M.; Fuso, A.; Pierandrei, S.; Bruno, S. M.; Testino, G.; Cimino, G.; Angeloni, A.; Lucarelli, M.. - In: DIAGNOSTICS. - ISSN 2075-4418. - 11:2(2021), p. 168. [10.3390/diagnostics11020168]

Quantitative evaluation of CFTR pre-mRNA splicing dependent on the (TG)mTn poly-variant tract

Sterrantino M.;Fuso A.;Pierandrei S.;Bruno S. M.;Testino G.;Angeloni A.;Lucarelli M.
2021

Abstract

Genetic analysis in cystic fibrosis (CF) is a difficult task. Within the many causes of variability and uncertainty, a major determinant is poor knowledge of the functional effect of most DNA variants of the Cystic Fibrosis Transmembrane conductance Regulator (CFTR) gene. In turn, knowledge of the effect of a CFTR variant has dramatic diagnostic, prognostic and, in the era of CF precision medicine, also therapeutic consequences. One of the most challenging CFTR variants is the (TG)mTn haplotype, which has variable functional effect and controversial clinical consequences. The exact quantification of the anomalous splicing of CFTR exon 10 (in the HGVS name; exon 9 in the legacy name) and, consequently, of the residual wild-type functional CFTR mRNA, should be mandatory in clinical assessment of patients with potentially pathological haplotype of this tract. Here, we present a real time-based assay for the quantification of the proportion of exon 10+/exon 10- CFTR mRNA, starting from nasal brushing. Our assay proved rapid, economic and easy to perform. Specific primers used for this assay are either disclosed or commercially available, allowing any laboratory to easily perform it. A simplified analysis of the data is provided, facilitating the interpretation of the results. This method helps to enhance the comprehension of the genotype- phenotype relationship in CF and CFTR-related disorders (CFTR-RD), crucial for the diagnosis, prognosis and personalized therapy of CF.
2021
(TG)mTn tract; CFTRrelated disorders (CFTR-RD); Cystic Fibrosis; Cystic Fibrosis Transmembrane conductance Regulator (CFTR); Functional effect of CFTR variants; Nasal brushing; Pre-mRNA splicing; Real time
01 Pubblicazione su rivista::01a Articolo in rivista
Quantitative evaluation of CFTR pre-mRNA splicing dependent on the (TG)mTn poly-variant tract / Sterrantino, M.; Fuso, A.; Pierandrei, S.; Bruno, S. M.; Testino, G.; Cimino, G.; Angeloni, A.; Lucarelli, M.. - In: DIAGNOSTICS. - ISSN 2075-4418. - 11:2(2021), p. 168. [10.3390/diagnostics11020168]
File allegati a questo prodotto
File Dimensione Formato  
Sterrantino_Quantitative-evaluation_2021.pdf

accesso aperto

Tipologia: Versione editoriale (versione pubblicata con il layout dell'editore)
Licenza: Tutti i diritti riservati (All rights reserved)
Dimensione 3.82 MB
Formato Adobe PDF
3.82 MB Adobe PDF

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1544437
Citazioni
  • ???jsp.display-item.citation.pmc??? 3
  • Scopus 5
  • ???jsp.display-item.citation.isi??? 5
social impact