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Exon Skipping in Directly Reprogrammed Myotubes Obtained from Human Urine-Derived Cells
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Genetics
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JoVE Journal Genetics
Exon Skipping in Directly Reprogrammed Myotubes Obtained from Human Urine-Derived Cells

Exon Skipping in Directly Reprogrammed Myotubes Obtained from Human Urine-Derived Cells

DOI:
10.3791/60840-v

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06:20 min

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May 07, 2020

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, ,
1Department of Molecular Therapy, National Institute of Neuroscience,National Center of Neurology and Psychiatry

Chapters

  • 00:04Introduction
  • 00:54Isolation and Primary Culture of Urine-Derived Cells
  • 02:31Infection with MYOD1-Retroviral Vector
  • 03:20Myogenic Differentiation of MYOD1-Transduced UDCs
  • 03:53Evaluation of Exon Skipping Efficiency by RT-PCR
  • 05:05Results: Restoration of Dystrophin mRNA in UDCs
  • 05:57Conclusion

Summary

Automatic Translation

Automatic Translation

In this article, we describe a detailed protocol for efficient modelling of Duchenne muscular dystrophy muscle using MYOD1-converted urine-derived cells to evaluate the restoration of dystrophin mRNA and protein levels after exon skipping.

Tags

Keywords: Exon SkippingDirect ReprogrammingHuman Urine-derived CellsMyotubesDuchenne Muscular Dystrophy3-deazaneplanocin A HydrochlorideDZnepHistone Methyltransferase InhibitorAutologousPrecision MedicineMuscular DiseasesCentrifugationCell CultureGelatin-coatedRetrovirusMYOD1Puromycin

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