Restoration of Dystrophin Expression in Mdx-Derived Muscle Progenitor Cells Using CRISPR/Cas9 System and Homology-Directed Repair Technology

Yue Jin, Yan Shen, Il man Kim, Neal L. Weintraub, Mark Hamrick, Yaoliang Tang

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Duchenne muscular dystrophy (DMD) is a progressive myopathy caused by mutations in genes encoding dystrophin proteins that ultimately lead to depletion of myogenic progenitor cells (MPCs). Several approaches have been used to correctly express the dystrophin gene in induced pluripotent stem cells (iPSCs), including deletion of mutated exon 23 (ΔEx23) by clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated gene 9 (Cas9)-mediated gene editing technology. However, this approach is labor-intensive due to individual colony picking and genotyping to verify allelic modification. Here, we present a protocol to restore the function of the dystrophin gene by using homology-directed repair (HDR)-based CRISPR/Cas9 and inducing myogenic program of reprogrammed iPSCs from Mdx mice by inducible muscle-specific transcription factor MyoD.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages455-464
Number of pages10
DOIs
StatePublished - 2023

Publication series

NameMethods in Molecular Biology
Volume2587
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • CRISPR-Cas9-HDR gene editing
  • Duchenne muscular dystrophy
  • iPSC-derived myoprogenitors

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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