Functional correction of dystrophin actin binding domain mutations by genome editing
| Autor: | Malte Tiburcy, John M. Shelton, Viktoriia Kyrychenko, Rhonda Bassel-Duby, Sergii Kyrychenko, Wolfram-Hubertus Zimmermann, Chengzu Long, Jay W. Schneider, Eric N. Olson |
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| Rok vydání: | 2017 |
| Předmět: |
musculoskeletal diseases
0301 basic medicine congenital hereditary and neonatal diseases and abnormalities Duchenne muscular dystrophy Induced Pluripotent Stem Cells Dystrophin 03 medical and health sciences Exon 0302 clinical medicine Protein Domains Genome editing Utrophin medicine Humans CRISPR Actin Gene Editing Genetics biology Exons General Medicine medicine.disease Actin cytoskeleton Actins Muscular Dystrophy Duchenne 030104 developmental biology Mutation biology.protein CRISPR-Cas Systems 030217 neurology & neurosurgery Protein Binding Research Article |
| Zdroj: | JCI Insight. 2 |
| ISSN: | 2379-3708 |
| DOI: | 10.1172/jci.insight.95918 |
| Popis: | Dystrophin maintains the integrity of striated muscles by linking the actin cytoskeleton with the cell membrane. Duchenne muscular dystrophy (DMD) is caused by mutations in the dystrophin gene (DMD) that result in progressive, debilitating muscle weakness, cardiomyopathy, and a shortened lifespan. Mutations of dystrophin that disrupt the amino-terminal actin-binding domain 1 (ABD-1), encoded by exons 2–8, represent the second-most common cause of DMD. In the present study, we compared three different strategies for CRISPR/Cas9 genome editing to correct mutations in the ABD-1 region of the DMD gene by deleting exons 3–9, 6–9, or 7–11 in human induced pluripotent stem cells (iPSCs) and by assessing the function of iPSC-derived cardiomyocytes. All three exon deletion strategies enabled the expression of truncated dystrophin protein and restoration of cardiomyocyte contractility and calcium transients to varying degrees. We show that deletion of exons 3–9 by genomic editing provides an especially effective means of correcting disease-causing ABD-1 mutations. These findings represent an important step toward eventual correction of common DMD mutations and provide a means of rapidly assessing the expression and function of internally truncated forms of dystrophin-lacking portions of ABD-1. |
| Databáze: | OpenAIRE |
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