1. Targeted addition of mini-dystrophin into rDNA locus of Duchenne muscular dystrophy patient-derived iPSCs.
- Author
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Zeng B, Zhou M, Liu B, Shen F, Xiao R, Su J, Hu Z, Zhang Y, Gu A, Wu L, Liu X, and Liang D
- Subjects
- Cell Differentiation, Cell Line, Cellular Reprogramming Techniques, Dystrophin metabolism, Exons, Gene Expression, Gene Targeting methods, Humans, Induced Pluripotent Stem Cells cytology, Loss of Function Mutation, Male, Muscular Dystrophy, Duchenne urine, Myocytes, Cardiac cytology, Myocytes, Cardiac metabolism, Proof of Concept Study, Recombinant Proteins genetics, Recombinant Proteins metabolism, Sequence Deletion, Urine cytology, DNA, Ribosomal genetics, Dystrophin genetics, Genetic Therapy methods, Induced Pluripotent Stem Cells metabolism, Muscular Dystrophy, Duchenne genetics, Muscular Dystrophy, Duchenne therapy
- Abstract
Duchenne muscular dystrophy (DMD), the most common lethal muscular disorder, affects 1 in 5000 male births. It is caused by mutations in the X-linked dystrophin gene (DMD), and there is no effective treatment currently. Gene addition is a promising strategy owing to its universality for patients with all gene mutations types. In this study, we describe a site-specific gene addition strategy in induced pluripotent stem cells (iPSCs) derived from a DMD patient with exon 50 deletion. By using transcription activator-like effector nickases (TALENickases), the mini-dystrophin cassette was precisely targeted at the ribosomal RNA gene (rDNA) locus via homologous recombination with high targeting efficiency. The targeted clone retained the main pluripotent properties and was differentiated into cardiomyocytes. Significantly, the dystrophin expression and membrane localization were restored in the genetic corrected iPSCs and their derived cardiomyocytes. More importantly, the enhanced spontaneous contraction was observed in modified cardiomyocytes. These results provide a proof of principle for an efficient targeted gene addition for DMD gene therapy and represents a significant step toward precisely therapeutic for DMD., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2021 Elsevier Inc. All rights reserved.)
- Published
- 2021
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