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In-Frame Deletion of Dystrophin Exons 8–50 Results in DMD Phenotype

Mutations that prevent the production of proteins in the DMD gene cause Duchenne muscular dystrophy. Most frequently, these are deletions leading to reading-frame shift. The “reading-frame rule” states that deletions that preserve ORF result in a milder Becker muscular dystrophy. By removing several...

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Autores principales: Egorova, Tatiana V., Galkin, Ivan I., Velyaev, Oleg A., Vassilieva, Svetlana G., Savchenko, Irina M., Loginov, Vyacheslav A., Dzhenkova, Marina A., Korshunova, Diana S., Kozlova, Olga S., Ivankov, Dmitry N., Polikarpova, Anna V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10252864/
https://www.ncbi.nlm.nih.gov/pubmed/37298068
http://dx.doi.org/10.3390/ijms24119117
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author Egorova, Tatiana V.
Galkin, Ivan I.
Velyaev, Oleg A.
Vassilieva, Svetlana G.
Savchenko, Irina M.
Loginov, Vyacheslav A.
Dzhenkova, Marina A.
Korshunova, Diana S.
Kozlova, Olga S.
Ivankov, Dmitry N.
Polikarpova, Anna V.
author_facet Egorova, Tatiana V.
Galkin, Ivan I.
Velyaev, Oleg A.
Vassilieva, Svetlana G.
Savchenko, Irina M.
Loginov, Vyacheslav A.
Dzhenkova, Marina A.
Korshunova, Diana S.
Kozlova, Olga S.
Ivankov, Dmitry N.
Polikarpova, Anna V.
author_sort Egorova, Tatiana V.
collection PubMed
description Mutations that prevent the production of proteins in the DMD gene cause Duchenne muscular dystrophy. Most frequently, these are deletions leading to reading-frame shift. The “reading-frame rule” states that deletions that preserve ORF result in a milder Becker muscular dystrophy. By removing several exons, new genome editing tools enable reading-frame restoration in DMD with the production of BMD-like dystrophins. However, not every truncated dystrophin with a significant internal loss functions properly. To determine the effectiveness of potential genome editing, each variant should be carefully studied in vitro or in vivo. In this study, we focused on the deletion of exons 8–50 as a potential reading-frame restoration option. Using the CRISPR-Cas9 tool, we created the novel mouse model DMDdel8-50, which has an in-frame deletion in the DMD gene. We compared DMDdel8-50 mice to C57Bl6/CBA background control mice and previously generated DMDdel8-34 KO mice. We discovered that the shortened protein was expressed and correctly localized on the sarcolemma. The truncated protein, on the other hand, was unable to function like a full-length dystrophin and prevent disease progression. On the basis of protein expression, histological examination, and physical assessment of the mice, we concluded that the deletion of exons 8–50 is an exception to the reading-frame rule.
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spelling pubmed-102528642023-06-10 In-Frame Deletion of Dystrophin Exons 8–50 Results in DMD Phenotype Egorova, Tatiana V. Galkin, Ivan I. Velyaev, Oleg A. Vassilieva, Svetlana G. Savchenko, Irina M. Loginov, Vyacheslav A. Dzhenkova, Marina A. Korshunova, Diana S. Kozlova, Olga S. Ivankov, Dmitry N. Polikarpova, Anna V. Int J Mol Sci Article Mutations that prevent the production of proteins in the DMD gene cause Duchenne muscular dystrophy. Most frequently, these are deletions leading to reading-frame shift. The “reading-frame rule” states that deletions that preserve ORF result in a milder Becker muscular dystrophy. By removing several exons, new genome editing tools enable reading-frame restoration in DMD with the production of BMD-like dystrophins. However, not every truncated dystrophin with a significant internal loss functions properly. To determine the effectiveness of potential genome editing, each variant should be carefully studied in vitro or in vivo. In this study, we focused on the deletion of exons 8–50 as a potential reading-frame restoration option. Using the CRISPR-Cas9 tool, we created the novel mouse model DMDdel8-50, which has an in-frame deletion in the DMD gene. We compared DMDdel8-50 mice to C57Bl6/CBA background control mice and previously generated DMDdel8-34 KO mice. We discovered that the shortened protein was expressed and correctly localized on the sarcolemma. The truncated protein, on the other hand, was unable to function like a full-length dystrophin and prevent disease progression. On the basis of protein expression, histological examination, and physical assessment of the mice, we concluded that the deletion of exons 8–50 is an exception to the reading-frame rule. MDPI 2023-05-23 /pmc/articles/PMC10252864/ /pubmed/37298068 http://dx.doi.org/10.3390/ijms24119117 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Egorova, Tatiana V.
Galkin, Ivan I.
Velyaev, Oleg A.
Vassilieva, Svetlana G.
Savchenko, Irina M.
Loginov, Vyacheslav A.
Dzhenkova, Marina A.
Korshunova, Diana S.
Kozlova, Olga S.
Ivankov, Dmitry N.
Polikarpova, Anna V.
In-Frame Deletion of Dystrophin Exons 8–50 Results in DMD Phenotype
title In-Frame Deletion of Dystrophin Exons 8–50 Results in DMD Phenotype
title_full In-Frame Deletion of Dystrophin Exons 8–50 Results in DMD Phenotype
title_fullStr In-Frame Deletion of Dystrophin Exons 8–50 Results in DMD Phenotype
title_full_unstemmed In-Frame Deletion of Dystrophin Exons 8–50 Results in DMD Phenotype
title_short In-Frame Deletion of Dystrophin Exons 8–50 Results in DMD Phenotype
title_sort in-frame deletion of dystrophin exons 8–50 results in dmd phenotype
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10252864/
https://www.ncbi.nlm.nih.gov/pubmed/37298068
http://dx.doi.org/10.3390/ijms24119117
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