From gRNA Identification to the Restoration of Dystrophin Expression: A Dystrophin Gene Correction Strategy for Duchenne Muscular Dystrophy Mutations Using the CRISPR-Induced Deletion Method

  • Benjamin Duchêne
  • Jean-Paul Iyombe-Engembe
  • Joël Rousseau
  • Jacques P. Tremblay
  • Dominique L. Ouellet
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1687)

Abstract

The discovery of the CRISPR-Cas9 system raises hope for the treatment of many genetic disorders. We describe here an approach based on the use of a pair of single guide RNAs to form a hybrid exon that does not only restore the dystrophin gene reading frame but also results in the production of a dystrophin protein with an adequate structure of the central rod-domain, with a correct spectrin-like repeat. The therapeutic approach described here involved DMD patient cells having a deletion of exons 51–53 of the DMD gene.

Key words

CRISPR-Cas9 Duchenne muscular dystrophy (DMD) Dystrophin Hybrid exon Spectrin-like repeats 

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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  • Benjamin Duchêne
    • 1
  • Jean-Paul Iyombe-Engembe
    • 1
  • Joël Rousseau
    • 1
  • Jacques P. Tremblay
    • 1
  • Dominique L. Ouellet
    • 1
  1. 1.Centre de Recherche du CHU de Québec-Université LavalQuébecCanada

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