Duchenne muscular dystrophy (DMD) is a genetic disorder that causes progressive muscle degeneration and weakness due to mutations in the dystrophin gene, which leads to the absence or malfunction of the dystrophin protein. “Protein replacement therapies”, which currently involve introducing functional but truncated dystrophin protein into muscle cells, are being explored as a potential treatment option for DMD. No attempts at full length DMD protein replacement have been attempted due to the size of the full length DMD protein.
We aimed to determine if it is possible to synthesize the full length dystrophin protein using a vector transfected suspension adapted Human Embryonic Kidney. We also sought to identify the concentration of DNA that would be most effective in producing high yields of the protein. We used various methods including enzyme-linked immunoassays (ELISA), Western blots, and flow cytometry to assess the effectiveness of plasmid-mediated dystrophin expression in FreeStyle 293 cells.
Based on our assay results, most specifically the Western blot, which showed a protein band at ~400kDa, it can be inferred that exogenous dystrophin expression is possible in FreeStyle 293 cells. Further research is needed to optimize conditions for the production and purification of high yields of pure dystrophin protein and to assess the safety and effectiveness of this approach in clinical trials.