Design of Liposome Formulations for CRISPR/Cas9 Enzyme Immobilization: Evaluation of 5-Alpha-Reductase Enzyme Knockout for Androgenic Disorders

The document discusses a study on a new treatment for androgen-related disorders such as prostate cancer and benign prostatic hyperplasia. The treatment uses liposome formulations to immobilize the CRISPR/Cas9 enzyme, which then deactivates the 5-alpha-reductase enzyme (SRD5α2) that converts testosterone to a potent androgen involved in these disorders. The liposome formulation was evaluated on various parameters and showed successful immobilization of the Cas9 protein. The formulation had an encapsulation efficiency of 80.60% and a loading capacity of 12.59% for the Cas9 enzyme. It demonstrated dose-dependent cytotoxicity on L929 mouse fibroblast cells and DU145 cells. The Lipo/Cas9:sgRNA formulation successfully edited genes, as shown by a 29.7% decrease in relative mRNA expression compared to the control. This approach could decrease the unwanted effects of DHT without changing the serum testosterone level, providing an alternative to 5-alpha-reductase enzyme inhibitors. However, challenges remain in directing therapeutic genes to target cells and ensuring higher gene knockout efficiency.