Gene Repair of iPSC Line with GARS (G294R) Mutation of CMT2D Disease by CRISPR/Cas9

Curr Med Sci. 2023 Apr;43(2):261-267. doi: 10.1007/s11596-023-2707-8. Epub 2023 Mar 18.


Objective: Charcot-Marie-Tooth disease (CMT) severely affects patient activity, and may cause disability. However, no clinical treatment is available to reverse the disease course. The combination of CRISPR/Cas9 and iPSCs may have therapeutic potential against nervous diseases, such as CMT.

Methods: In the present study, the skin fibroblasts of CMT type 2D (CMT2D) patients with the c.880G>A heterozygous nucleotide mutation in the GARS gene were reprogrammed into iPSCs using three plasmids (pCXLE-hSK, pCXLE-hUL and pCXLE-hOCT3/4-shp5-F). Then, CRISPR/Cas9 technology was used to repair the mutated gene sites at the iPSC level.

Results: An iPSC line derived from the GARS (G294R) family with fibular atrophy was successfully induced, and the mutated gene loci were repaired at the iPSC level using CRISPR/Cas9 technology. These findings lay the foundation for future research on drug screening and cell therapy.

Conclusion: iPSCs can differentiate into different cell types, and originate from autologous cells. Therefore, they are promising for the development of autologous cell therapies for degenerative diseases. The combination of CRISPR/Cas9 and iPSCs may open a new avenue for the treatment of nervous diseases, such as CMT.

Keywords: CRISPR/Cas9; Charcot-Marie-Tooth disease; GARS; gene therapy; iPSCs.

MeSH terms

  • CRISPR-Cas Systems / genetics
  • Charcot-Marie-Tooth Disease* / genetics
  • Charcot-Marie-Tooth Disease* / metabolism
  • Charcot-Marie-Tooth Disease* / therapy
  • Humans
  • Induced Pluripotent Stem Cells* / metabolism
  • Mutation
  • Targeted Gene Repair* / methods

Supplementary concepts

  • Charcot-Marie-Tooth disease, Type 2D