Significance
The goal of this project was to develop consistently high and reliable expression of therapeutic genes in mammalian cells, which is one of the key challenges for gene therapy. The present work contributes directly to the development of safer and more effective gene treatments by evaluating the impact of different codon optimization strategies on protein output for the relevant genes, namely ST3GAL5, SMN1, and A1AT. An increase in expression of therapeutic genes strengthens their effects and allows for a potential decrease in vector dosages, enhancing long-term gene stability, and even possible cures for diseases. Even though the optimized A1AT constructs showed better protein expression, the lack of clear improvement for ST3GAL5 and SMN1 really shows the need to pick suitable cell lines and have reliable detection methods when testing for gene expression. By identifying both the successes and limitations of our optimization model, this work offers valuable insights into how these strategies should be applied and tested in the future development of therapies. For those reasons, this research is highly relevant to the further development of gene therapy technologies.
