Abstract
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.
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Conflict of Interest Statement
The authors declare that they have no conflicts of interest.
This work was supported by grants from the National Major Scientific and Technological Special Project for “Significant New Drugs Development” (No. 2019ZX09301159), the “Thousand Talent Program” for Science and Technology Innovation Leader in Henan (No. 194200510002), the Bingtuan Science and Technology Project (No. 2019AB034), and the Natural Science Foundation of Henan Province of China (No. 202300410381).
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Lu, Pj., Zhang, P., Liu, Yc. et al. Gene Repair of iPSC Line with GARS (G294R) Mutation of CMT2D Disease by CRISPR/Cas9. CURR MED SCI 43, 261–267 (2023). https://doi.org/10.1007/s11596-023-2707-8
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DOI: https://doi.org/10.1007/s11596-023-2707-8