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Publication details

Document type
Journal articles

Document subtype
Full paper

Title
Correction of a Cystic Fibrosis Splicing Mutation by Antisense Oligonucleotides

Participants in the publication
Susana Igreja (Author)
Luka A. Clarke (Author)
BioISI
 Hugo M. Botelho (Author)
BioISI - Biosystems & Integrative Sciences Institute
 Dep. Química e Bioquímica
 Luís Marques (Author)
Margarida D. Amaral (Author)
Dep. Química e Bioquímica
 BioISI
Summary
Cystic fibrosis (CF), the most common life-threatening genetic disease in Caucasians, is caused by ∼2,000 different mutations in the CF transmembrane conductance regulator (CFTR) gene. A significant fraction of these (∼13%) affect pre-mRNA splicing for which novel therapies have been somewhat neglected. We have previously described the effect of the CFTR splicing mutation c.2657+5G>A in IVS16, showing that it originates transcripts lacking exon 16 as well as wild-type transcripts. Here, we tested an RNA-based antisense oligonucleotide (AON) strategy to correct the aberrant splicing caused by this mutation. Two AONs (AON1/2) complementary to the pre-mRNA IVS16 mutant region were designed and their effect on splicing was assessed at the RNA and protein levels, on intracellular protein localization and function. To this end, we used the 2657+5G>A mutant CFTR minigene stably expressed in HEK293 Flp-In cells that express a single copy of the transgene. RNA data from AON1-treated mutant cells show that exon 16 inclusion was almost completely restored (to 95%), also resulting in increased levels of correctly localized CFTR protein at the plasma membrane (PM) and with increased function. A novel two-color CFTR splicing reporter minigene developed here allowed the quantitative monitoring of splicing by automated microscopy localization of CFTR at the PM. The AON strategy is thus a promising therapeutic approach for the specific correction of alternative splicing.

Date of Publication
2016

Institution
BioISI - Biosystems & Integrative Sciences Institute

Where published
Human Mutation

Publication Identifiers
ISSN - 1059-7794

Publisher
Wiley

Volume
37
Number
2

Starting page
209
Last page
215

Document Identifiers
URL - http://dx.doi.org/10.1002/humu.22931
DOI - https://doi.org/10.1002/humu.22931

Rankings
SCOPUS Q1 (2016) - 3.231 - Genetics
SCIMAGO Q1 (2016) - 3.231 - Genetics (clinical)
SCIMAGO Q1 (2016) - 3.231 - Genetics
SCOPUS Q1 (2016) - 3.231 - Genetics(clinical)


Export

APA
Susana Igreja, Luka A. Clarke, Hugo M. Botelho, Luís Marques, Margarida D. Amaral, (2016). Correction of a Cystic Fibrosis Splicing Mutation by Antisense Oligonucleotides. Human Mutation, 37, 209-215. ISSN 1059-7794. eISSN . http://dx.doi.org/10.1002/humu.22931

IEEE
Susana Igreja, Luka A. Clarke, Hugo M. Botelho, Luís Marques, Margarida D. Amaral, "Correction of a Cystic Fibrosis Splicing Mutation by Antisense Oligonucleotides" in Human Mutation, vol. 37, pp. 209-215, 2016. 10.1002/humu.22931

BIBTEX
@article{38920, author = {Susana Igreja and Luka A. Clarke and Hugo M. Botelho and Luís Marques and Margarida D. Amaral}, title = {Correction of a Cystic Fibrosis Splicing Mutation by Antisense Oligonucleotides}, journal = {Human Mutation}, year = 2016, pages = {209-215}, volume = 37 }