A Single CRISPR-Cas9 Deletion Strategy that Targets the Majority of DMD Patients Restores Dystrophin Function in hiPSC-Derived Muscle Cells
| Autor: | Young, Courtney S, Hicks, Michael R, Ermolova, Natalia V, Nakano, Haruko, Jan, Majib, Younesi, Shahab, Karumbayaram, Saravanan, Kumagai-Cresse, Chino, Wang, Derek, Zack, Jerome A, Kohn, Donald B, Nakano, Atsushi, Nelson, Stanley F, Miceli, M Carrie, Spencer, Melissa J, Pyle, April D |
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| Rok vydání: | 2016 |
| Předmět: |
Duchenne/ Becker Muscular Dystrophy
Intellectual and Developmental Disabilities (IDD) Induced Pluripotent Stem Cells SCID Regenerative Medicine Medical and Health Sciences Dystrophin Mice Rare Diseases Clinical Research Genetics Animals Humans 2.1 Biological and endogenous factors Muscular Dystrophy Aetiology Gene Editing Pediatric Stem Cell Research - Induced Pluripotent Stem Cell Neurosciences Skeletal Biological Sciences Duchenne Stem Cell Research Brain Disorders Musculoskeletal Muscle CRISPR-Cas Systems Gene Deletion Developmental Biology |
| Zdroj: | Cell stem cell, vol 18, iss 4 |
| Popis: | Mutations in DMD disrupt the reading frame, prevent dystrophin translation, and cause Duchenne muscular dystrophy (DMD). Here we describe a CRISPR/Cas9 platform applicable to 60% of DMD patient mutations. We applied the platform to DMD-derived hiPSCs where successful deletion and non-homologous end joining of up to 725 kb reframed the DMD gene. This is the largest CRISPR/Cas9-mediated deletion shown to date in DMD. Use of hiPSCs allowed evaluation of dystrophin in disease-relevant cell types. Cardiomyocytes and skeletal muscle myotubes derived from reframed hiPSC clonal lines had restored dystrophin protein. The internally deleted dystrophin was functional as demonstrated by improved membrane integrity and restoration of the dystrophin glycoprotein complex in vitro and in vivo. Furthermore, miR31 was reduced upon reframing, similar to observations in Becker muscular dystrophy. This work demonstrates the feasibility of using a single CRISPR pair to correct the reading frame for the majority of DMD patients. |
| Databáze: | OpenAIRE |
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