Background <p>Myoferlin (MYOF), a key gene in endocytosis, is known to be crucial for metastasis in cancer. While the involvement of MYOF as a biomarker is documented in cancer, a comprehensive study on its knockdown for targeted therapy via folate is a novel approach.</p> Methods <p>In the current study, the expression, pathways, mutations, and interacting genes of MYOF were analyzed via bioinformatics analysis, which included computational drug discovery and spatial transcriptomics coupled with generative artificial intelligence experiments that were performed to elucidate the role of folate in MYOF knockdown.</p> Results <p>The findings identified MYOF as a predominantly expressed gene biomarker in brain cancer genesis and its progression. Literature-reported mutations of MYOF were linked to pathways associated with depression, reinforcing the connection between brain cancer, tyrosine mutations, and depression in Caucasian patients. Additionally, computational drug discovery and spatial transcriptomics involving generative artificial intelligence experiments demonstrated folate as a potential therapeutic in MYOF knockdown.</p> Conclusions <p>These results suggest that dysregulation and mutation of MYOF position it as an oncogenic factor, and it serves as a potential therapeutic target for folate therapy for brain cancer associated with Caucasians.</p>