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A TALEN-Exon Skipping Design for a Bethlem Myopathy Model in Zebrafish

Presently, human collagen VI-related diseases such as Ullrich congenital muscular dystrophy (UCMD) and Bethlem myopathy (BM) remain incurable, emphasizing the need to unravel their etiology and improve their treatments. In UCMD, symptom onset occurs early, and both diseases aggravate with ageing. In...

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Autores principales: Radev, Zlatko, Hermel, Jean-Michel, Elipot, Yannick, Bretaud, Sandrine, Arnould, Sylvain, Duchateau, Philippe, Ruggiero, Florence, Joly, Jean-Stéphane, Sohm, Frédéric
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4519248/
https://www.ncbi.nlm.nih.gov/pubmed/26221953
http://dx.doi.org/10.1371/journal.pone.0133986
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author Radev, Zlatko
Hermel, Jean-Michel
Elipot, Yannick
Bretaud, Sandrine
Arnould, Sylvain
Duchateau, Philippe
Ruggiero, Florence
Joly, Jean-Stéphane
Sohm, Frédéric
author_facet Radev, Zlatko
Hermel, Jean-Michel
Elipot, Yannick
Bretaud, Sandrine
Arnould, Sylvain
Duchateau, Philippe
Ruggiero, Florence
Joly, Jean-Stéphane
Sohm, Frédéric
author_sort Radev, Zlatko
collection PubMed
description Presently, human collagen VI-related diseases such as Ullrich congenital muscular dystrophy (UCMD) and Bethlem myopathy (BM) remain incurable, emphasizing the need to unravel their etiology and improve their treatments. In UCMD, symptom onset occurs early, and both diseases aggravate with ageing. In zebrafish fry, morpholinos reproduced early UCMD and BM symptoms but did not allow to study the late phenotype. Here, we produced the first zebrafish line with the human mutation frequently found in collagen VI-related disorders such as UCMD and BM. We used a transcription activator-like effector nuclease (TALEN) to design the col6a1(ama605003)-line with a mutation within an essential splice donor site, in intron 14 of the col6a1 gene, which provoke an in-frame skipping of exon 14 in the processed mRNA. This mutation at a splice donor site is the first example of a template-independent modification of splicing induced in zebrafish using a targetable nuclease. This technique is readily expandable to other organisms and can be instrumental in other disease studies. Histological and ultrastructural analyzes of homozygous and heterozygous mutant fry and 3 months post-fertilization (mpf) fish revealed co-dominantly inherited abnormal myofibers with disorganized myofibrils, enlarged sarcoplasmic reticulum, altered mitochondria and misaligned sarcomeres. Locomotion analyzes showed hypoxia-response behavior in 9 mpf col6a1 mutant unseen in 3 mpf fish. These symptoms worsened with ageing as described in patients with collagen VI deficiency. Thus, the col6a1(ama605003)-line is the first adult zebrafish model of collagen VI-related diseases; it will be instrumental both for basic research and drug discovery assays focusing on this type of disorders.
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spelling pubmed-45192482015-07-31 A TALEN-Exon Skipping Design for a Bethlem Myopathy Model in Zebrafish Radev, Zlatko Hermel, Jean-Michel Elipot, Yannick Bretaud, Sandrine Arnould, Sylvain Duchateau, Philippe Ruggiero, Florence Joly, Jean-Stéphane Sohm, Frédéric PLoS One Research Article Presently, human collagen VI-related diseases such as Ullrich congenital muscular dystrophy (UCMD) and Bethlem myopathy (BM) remain incurable, emphasizing the need to unravel their etiology and improve their treatments. In UCMD, symptom onset occurs early, and both diseases aggravate with ageing. In zebrafish fry, morpholinos reproduced early UCMD and BM symptoms but did not allow to study the late phenotype. Here, we produced the first zebrafish line with the human mutation frequently found in collagen VI-related disorders such as UCMD and BM. We used a transcription activator-like effector nuclease (TALEN) to design the col6a1(ama605003)-line with a mutation within an essential splice donor site, in intron 14 of the col6a1 gene, which provoke an in-frame skipping of exon 14 in the processed mRNA. This mutation at a splice donor site is the first example of a template-independent modification of splicing induced in zebrafish using a targetable nuclease. This technique is readily expandable to other organisms and can be instrumental in other disease studies. Histological and ultrastructural analyzes of homozygous and heterozygous mutant fry and 3 months post-fertilization (mpf) fish revealed co-dominantly inherited abnormal myofibers with disorganized myofibrils, enlarged sarcoplasmic reticulum, altered mitochondria and misaligned sarcomeres. Locomotion analyzes showed hypoxia-response behavior in 9 mpf col6a1 mutant unseen in 3 mpf fish. These symptoms worsened with ageing as described in patients with collagen VI deficiency. Thus, the col6a1(ama605003)-line is the first adult zebrafish model of collagen VI-related diseases; it will be instrumental both for basic research and drug discovery assays focusing on this type of disorders. Public Library of Science 2015-07-29 /pmc/articles/PMC4519248/ /pubmed/26221953 http://dx.doi.org/10.1371/journal.pone.0133986 Text en © 2015 Radev et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Radev, Zlatko
Hermel, Jean-Michel
Elipot, Yannick
Bretaud, Sandrine
Arnould, Sylvain
Duchateau, Philippe
Ruggiero, Florence
Joly, Jean-Stéphane
Sohm, Frédéric
A TALEN-Exon Skipping Design for a Bethlem Myopathy Model in Zebrafish
title A TALEN-Exon Skipping Design for a Bethlem Myopathy Model in Zebrafish
title_full A TALEN-Exon Skipping Design for a Bethlem Myopathy Model in Zebrafish
title_fullStr A TALEN-Exon Skipping Design for a Bethlem Myopathy Model in Zebrafish
title_full_unstemmed A TALEN-Exon Skipping Design for a Bethlem Myopathy Model in Zebrafish
title_short A TALEN-Exon Skipping Design for a Bethlem Myopathy Model in Zebrafish
title_sort talen-exon skipping design for a bethlem myopathy model in zebrafish
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4519248/
https://www.ncbi.nlm.nih.gov/pubmed/26221953
http://dx.doi.org/10.1371/journal.pone.0133986
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