Targeted addition of mini-dystrophin into rDNA locus of Duchenne muscular dystrophy patient-derived iPSCs
Autor: | Desheng Liang, Rou Xiao, Bo Liu, Xionghao Liu, Lingqian Wu, Jiasun Su, Yiti Zhang, Baitao Zeng, Ao Gu, Zhiqing Hu, Fei Shen, Miaojin Zhou |
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Rok vydání: | 2021 |
Předmět: |
Male
musculoskeletal diseases 0301 basic medicine congenital hereditary and neonatal diseases and abnormalities Duchenne muscular dystrophy Induced Pluripotent Stem Cells Biophysics Clone (cell biology) Gene Expression Locus (genetics) Urine Gene mutation Biology DNA Ribosomal Proof of Concept Study Biochemistry Cell Line Dystrophin 03 medical and health sciences Exon 0302 clinical medicine Loss of Function Mutation medicine Humans Cellular Reprogramming Techniques Myocytes Cardiac Induced pluripotent stem cell Molecular Biology Gene Sequence Deletion Cell Differentiation Exons Genetic Therapy Cell Biology medicine.disease Recombinant Proteins Cell biology Muscular Dystrophy Duchenne 030104 developmental biology 030220 oncology & carcinogenesis Gene Targeting biology.protein |
Zdroj: | Biochemical and Biophysical Research Communications. 545:40-45 |
ISSN: | 0006-291X |
Popis: | 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. |
Databáze: | OpenAIRE |
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