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CRISPR/Cas9-generated mouse model of Duchenne muscular dystrophy recapitulating a newly identified large 430 kb deletion in the human DMD gene
Exon skipping is a promising strategy for Duchenne muscular dystrophy (DMD) disease-modifying therapy. To make this approach safe, ensuring that excluding one or more exons will restore the reading frame and that the resulting protein will retain critical functions of the full-length dystrophin prot...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Company of Biologists Ltd
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6505476/ https://www.ncbi.nlm.nih.gov/pubmed/31028078 http://dx.doi.org/10.1242/dmm.037655 |
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author | Egorova, Tatiana V. Zotova, Evgenia D. Reshetov, Denis A. Polikarpova, Anna V. Vassilieva, Svetlana G. Vlodavets, Dmitry V. Gavrilov, Alexey A. Ulianov, Sergey V. Buchman, Vladimir L. Deykin, Alexei V. |
author_facet | Egorova, Tatiana V. Zotova, Evgenia D. Reshetov, Denis A. Polikarpova, Anna V. Vassilieva, Svetlana G. Vlodavets, Dmitry V. Gavrilov, Alexey A. Ulianov, Sergey V. Buchman, Vladimir L. Deykin, Alexei V. |
author_sort | Egorova, Tatiana V. |
collection | PubMed |
description | Exon skipping is a promising strategy for Duchenne muscular dystrophy (DMD) disease-modifying therapy. To make this approach safe, ensuring that excluding one or more exons will restore the reading frame and that the resulting protein will retain critical functions of the full-length dystrophin protein is necessary. However, in vivo testing of the consequences of skipping exons that encode the N-terminal actin-binding domain (ABD) has been confounded by the absence of a relevant animal model. We created a mouse model of the disease recapitulating a novel human mutation, a large de novo deletion of exons 8-34 of the DMD gene, found in a Russian DMD patient. This mutation was achieved by deleting exons 8-34 of the X-linked mouse Dmd gene using CRISPR/Cas9 genome editing, which led to a reading frame shift and the absence of functional dystrophin production. Male mice carrying this deletion display several important signs of muscular dystrophy, including a gradual age-dependent decrease in muscle strength, increased creatine kinase, muscle fibrosis and central nucleation. The degrees of these changes are comparable to those observed in mdx mice, a standard laboratory model of DMD. This new model of DMD will be useful for validating therapies based on skipping exons that encode the N-terminal ABD and for improving our understanding of the role of the N-terminal domain and central rod domain in the biological function of dystrophin. Simultaneous skipping of exons 6 and 7 should restore the gene reading frame and lead to the production of a protein that might retain functionality despite the partial deletion of the ABD. |
format | Online Article Text |
id | pubmed-6505476 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | The Company of Biologists Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-65054762019-05-09 CRISPR/Cas9-generated mouse model of Duchenne muscular dystrophy recapitulating a newly identified large 430 kb deletion in the human DMD gene Egorova, Tatiana V. Zotova, Evgenia D. Reshetov, Denis A. Polikarpova, Anna V. Vassilieva, Svetlana G. Vlodavets, Dmitry V. Gavrilov, Alexey A. Ulianov, Sergey V. Buchman, Vladimir L. Deykin, Alexei V. Dis Model Mech Resource Article Exon skipping is a promising strategy for Duchenne muscular dystrophy (DMD) disease-modifying therapy. To make this approach safe, ensuring that excluding one or more exons will restore the reading frame and that the resulting protein will retain critical functions of the full-length dystrophin protein is necessary. However, in vivo testing of the consequences of skipping exons that encode the N-terminal actin-binding domain (ABD) has been confounded by the absence of a relevant animal model. We created a mouse model of the disease recapitulating a novel human mutation, a large de novo deletion of exons 8-34 of the DMD gene, found in a Russian DMD patient. This mutation was achieved by deleting exons 8-34 of the X-linked mouse Dmd gene using CRISPR/Cas9 genome editing, which led to a reading frame shift and the absence of functional dystrophin production. Male mice carrying this deletion display several important signs of muscular dystrophy, including a gradual age-dependent decrease in muscle strength, increased creatine kinase, muscle fibrosis and central nucleation. The degrees of these changes are comparable to those observed in mdx mice, a standard laboratory model of DMD. This new model of DMD will be useful for validating therapies based on skipping exons that encode the N-terminal ABD and for improving our understanding of the role of the N-terminal domain and central rod domain in the biological function of dystrophin. Simultaneous skipping of exons 6 and 7 should restore the gene reading frame and lead to the production of a protein that might retain functionality despite the partial deletion of the ABD. The Company of Biologists Ltd 2019-04-01 2019-04-25 /pmc/articles/PMC6505476/ /pubmed/31028078 http://dx.doi.org/10.1242/dmm.037655 Text en © 2019. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by/4.0This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed. |
spellingShingle | Resource Article Egorova, Tatiana V. Zotova, Evgenia D. Reshetov, Denis A. Polikarpova, Anna V. Vassilieva, Svetlana G. Vlodavets, Dmitry V. Gavrilov, Alexey A. Ulianov, Sergey V. Buchman, Vladimir L. Deykin, Alexei V. CRISPR/Cas9-generated mouse model of Duchenne muscular dystrophy recapitulating a newly identified large 430 kb deletion in the human DMD gene |
title | CRISPR/Cas9-generated mouse model of Duchenne muscular dystrophy recapitulating a newly identified large 430 kb deletion in the human DMD gene |
title_full | CRISPR/Cas9-generated mouse model of Duchenne muscular dystrophy recapitulating a newly identified large 430 kb deletion in the human DMD gene |
title_fullStr | CRISPR/Cas9-generated mouse model of Duchenne muscular dystrophy recapitulating a newly identified large 430 kb deletion in the human DMD gene |
title_full_unstemmed | CRISPR/Cas9-generated mouse model of Duchenne muscular dystrophy recapitulating a newly identified large 430 kb deletion in the human DMD gene |
title_short | CRISPR/Cas9-generated mouse model of Duchenne muscular dystrophy recapitulating a newly identified large 430 kb deletion in the human DMD gene |
title_sort | crispr/cas9-generated mouse model of duchenne muscular dystrophy recapitulating a newly identified large 430 kb deletion in the human dmd gene |
topic | Resource Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6505476/ https://www.ncbi.nlm.nih.gov/pubmed/31028078 http://dx.doi.org/10.1242/dmm.037655 |
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