Correction of exon 2, exon 2-9 and exons 8-9 duplications in DMD patient myogenic cells by a single CRISPR/Cas9 system.
Juliette LemoineAuriane DuboisAlan DorvalAbbass JaberGanesh WarthiKamel MamchaouiTao WangGuillaume CorreMatteo BovolentaIsabelle RichardPublished in: Scientific reports (2024)
Duchenne Muscular dystrophy (DMD), a yet-incurable X-linked recessive disorder that results in muscle wasting and loss of ambulation is due to mutations in the dystrophin gene. Exonic duplications of dystrophin gene are a common type of mutations found in DMD patients. In this study, we utilized a single guide RNA CRISPR strategy targeting intronic regions to delete the extra duplicated regions in patient myogenic cells carrying duplication of exon 2, exons 2-9, and exons 8-9 in the DMD gene. Immunostaining on CRISPR-corrected derived myotubes demonstrated the rescue of dystrophin protein. Subsequent RNA sequencing of the DMD cells indicated rescue of genes of dystrophin related pathways. Examination of predicted close-match off-targets evidenced no aberrant gene editing at these loci. Here, we further demonstrate the efficiency of a single guide CRISPR strategy capable of deleting multi-exon duplications in the DMD gene without significant off target effect. Our study contributes valuable insights into the safety and efficacy of using single guide CRISPR strategy as a potential therapeutic approach for DMD patients with duplications of variable size.
Keyphrases
- duchenne muscular dystrophy
- genome wide
- crispr cas
- genome editing
- muscular dystrophy
- induced apoptosis
- dna methylation
- copy number
- cell cycle arrest
- genome wide identification
- skeletal muscle
- end stage renal disease
- endoplasmic reticulum stress
- case report
- single cell
- chronic kidney disease
- cell death
- gene expression
- prognostic factors
- cell proliferation
- small molecule
- autism spectrum disorder
- protein protein
- peritoneal dialysis