Gene-editing in patient and humanized-mice primary muscle stem cells rescues dysferlin expression in dysferlin-deficient muscular dystrophy

Escobar, H.; Di Francescantonio, S.; Smirnova, J.; Graf, R.; Müthel, S.; Marg, A.; Zhogov, A.; Krishna, S.; Metzler, E.; Petkova, M.; Daumke, O.; Kühn, R.; Spuler, S.

Gene-editing in patient and humanized-mice primary muscle stem cells rescues dysferlin expression in dysferlin-deficient muscular dystrophy

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Item Type:	Article
Title:	Gene-editing in patient and humanized-mice primary muscle stem cells rescues dysferlin expression in dysferlin-deficient muscular dystrophy
Creators Name:	Escobar, H., Di Francescantonio, S., Smirnova, J., Graf, R., Müthel, S., Marg, A., Zhogov, A., Krishna, S., Metzler, E., Petkova, M., Daumke, O., Kühn, R. and Spuler, S.
Abstract:	Dystrophy-associated fer-1-like protein (dysferlin) conducts plasma membrane repair. Mutations in the DYSF gene cause a panoply of genetic muscular dystrophies. We targeted a frequent loss-of-function, DYSF exon 44, founder frameshift mutation with mRNA-mediated delivery of SpCas9 in combination with a mutation-specific sgRNA to primary muscle stem cells from two homozygous patients. We observed a consistent >60% exon 44 re-framing, rescuing a full-length and functional dysferlin protein. A new mouse model harboring a humanized Dysf exon 44 with the founder mutation, hEx44mut, recapitulates the patients’ phenotype and an identical re-framing outcome in primary muscle stem cells. Finally, gene-edited murine primary muscle stem-cells are able to regenerate muscle and rescue dysferlin when transplanted back into hEx44mut hosts. These findings are the first to show that a CRISPR-mediated therapy can ameliorate dysferlin deficiency. We suggest that gene-edited primary muscle stem cells could exhibit utility, not only in treating dysferlin deficiency syndromes, but also perhaps other forms of muscular dystrophy.
Keywords:	Animal Disease Models, CRISPR-Cas Systems, Dysferlin, Exons, Frameshift Mutation, Gene Editing, Genetic Therapy, Limb-Girdle Muscular Dystrophies, Membrane Proteins, Muscular Dystrophies, Skeletal Muscle, Stem Cells, Animals, Mice
Source:	Nature Communications
ISSN:	2041-1723
Publisher:	Nature Publishing Group
Volume:	16
Number:	1
Page Range:	120
Date:	2 January 2025
Official Publication:	https://doi.org/10.1038/s41467-024-55086-0
PubMed:	View item in PubMed

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