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Sheep models of F508del and G542X cystic fibrosis mutations show cellular responses to human therapeutics
Cystic Fibrosis (CF) is a genetic disease caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. The F508del and G542X are the most common mutations found in US patients, accounting for 86.4% and 4.6% of all mutations, respectively. The F508del causes deletion of the phenylal...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8493969/ https://www.ncbi.nlm.nih.gov/pubmed/34632318 http://dx.doi.org/10.1096/fba.2021-00043 |
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author | Viotti Perisse, Iuri Fan, Zhiqiang Van Wettere, Arnaud Liu, Ying Leir, Shih‐Hsing Keim, Jacob Regouski, Misha Wilson, Michael D. Cholewa, Kelly M. Mansbach, Sara N. Kelley, Thomas J. Wang, Zhongde Harris, Ann White, Kenneth L. Polejaeva, Irina A. |
author_facet | Viotti Perisse, Iuri Fan, Zhiqiang Van Wettere, Arnaud Liu, Ying Leir, Shih‐Hsing Keim, Jacob Regouski, Misha Wilson, Michael D. Cholewa, Kelly M. Mansbach, Sara N. Kelley, Thomas J. Wang, Zhongde Harris, Ann White, Kenneth L. Polejaeva, Irina A. |
author_sort | Viotti Perisse, Iuri |
collection | PubMed |
description | Cystic Fibrosis (CF) is a genetic disease caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. The F508del and G542X are the most common mutations found in US patients, accounting for 86.4% and 4.6% of all mutations, respectively. The F508del causes deletion of the phenylalanine residue at position 508 and is associated with impaired CFTR protein folding. The G542X is a nonsense mutation that introduces a stop codon into the mRNA, thus preventing normal CFTR protein synthesis. Here, we describe the generation of CFTR(F508del) (/) (F508del) and CFTR(G542X) (/) (G542X) lambs using CRISPR/Cas9 and somatic cell nuclear transfer (SCNT). First, we introduced either F508del or G542X mutations into sheep fetal fibroblasts that were subsequently used as nuclear donors for SCNT. The newborn CF lambs develop pathology similar to CFTR (−/−) sheep and CF patients. Moreover, tracheal epithelial cells from the CFTR(F508del) (/) (F508del) lambs responded to a human CFTR (hCFTR) potentiator and correctors, and those from CFTR(G542X) (/) (G542X) lambs showed modest restoration of CFTR function following inhibition of nonsense‐mediated decay (NMD) and aminoglycoside antibiotic treatments. Thus, the phenotype and electrophysiology of these novel models represent an important advance for testing new CF therapeutics and gene therapy to improve the health of patients with this life‐limiting disorder. |
format | Online Article Text |
id | pubmed-8493969 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-84939692021-10-08 Sheep models of F508del and G542X cystic fibrosis mutations show cellular responses to human therapeutics Viotti Perisse, Iuri Fan, Zhiqiang Van Wettere, Arnaud Liu, Ying Leir, Shih‐Hsing Keim, Jacob Regouski, Misha Wilson, Michael D. Cholewa, Kelly M. Mansbach, Sara N. Kelley, Thomas J. Wang, Zhongde Harris, Ann White, Kenneth L. Polejaeva, Irina A. FASEB Bioadv Research Articles Cystic Fibrosis (CF) is a genetic disease caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. The F508del and G542X are the most common mutations found in US patients, accounting for 86.4% and 4.6% of all mutations, respectively. The F508del causes deletion of the phenylalanine residue at position 508 and is associated with impaired CFTR protein folding. The G542X is a nonsense mutation that introduces a stop codon into the mRNA, thus preventing normal CFTR protein synthesis. Here, we describe the generation of CFTR(F508del) (/) (F508del) and CFTR(G542X) (/) (G542X) lambs using CRISPR/Cas9 and somatic cell nuclear transfer (SCNT). First, we introduced either F508del or G542X mutations into sheep fetal fibroblasts that were subsequently used as nuclear donors for SCNT. The newborn CF lambs develop pathology similar to CFTR (−/−) sheep and CF patients. Moreover, tracheal epithelial cells from the CFTR(F508del) (/) (F508del) lambs responded to a human CFTR (hCFTR) potentiator and correctors, and those from CFTR(G542X) (/) (G542X) lambs showed modest restoration of CFTR function following inhibition of nonsense‐mediated decay (NMD) and aminoglycoside antibiotic treatments. Thus, the phenotype and electrophysiology of these novel models represent an important advance for testing new CF therapeutics and gene therapy to improve the health of patients with this life‐limiting disorder. John Wiley and Sons Inc. 2021-08-02 /pmc/articles/PMC8493969/ /pubmed/34632318 http://dx.doi.org/10.1096/fba.2021-00043 Text en © 2021 The Authors. FASEB BioAdvances published by Wiley Periodicals LLC on behalf of The Federation of American Societies for Experimental Biology https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Research Articles Viotti Perisse, Iuri Fan, Zhiqiang Van Wettere, Arnaud Liu, Ying Leir, Shih‐Hsing Keim, Jacob Regouski, Misha Wilson, Michael D. Cholewa, Kelly M. Mansbach, Sara N. Kelley, Thomas J. Wang, Zhongde Harris, Ann White, Kenneth L. Polejaeva, Irina A. Sheep models of F508del and G542X cystic fibrosis mutations show cellular responses to human therapeutics |
title | Sheep models of F508del and G542X cystic fibrosis mutations show cellular responses to human therapeutics |
title_full | Sheep models of F508del and G542X cystic fibrosis mutations show cellular responses to human therapeutics |
title_fullStr | Sheep models of F508del and G542X cystic fibrosis mutations show cellular responses to human therapeutics |
title_full_unstemmed | Sheep models of F508del and G542X cystic fibrosis mutations show cellular responses to human therapeutics |
title_short | Sheep models of F508del and G542X cystic fibrosis mutations show cellular responses to human therapeutics |
title_sort | sheep models of f508del and g542x cystic fibrosis mutations show cellular responses to human therapeutics |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8493969/ https://www.ncbi.nlm.nih.gov/pubmed/34632318 http://dx.doi.org/10.1096/fba.2021-00043 |
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