Vesicular stomatitis virus (VSV), a negative-sense RNA virus, has emerged as a versatile platform for oncolytic virotherapy and vaccine development. Numerous advantages, such as a broad host range, rapid replication, and simple and easily modifiable genomes, make it an attractive candidate for both the treatment and prevention of infectious diseases and tumors. Advances in molecular biology techniques, particularly reverse genetics and synthetic biology, have facilitated the precise engineering of VSV genomes, thereby significantly expanding their applications in cancer therapeutics. One of the most common antitumor strategies is to promote tumor cell death. Numerous studies have shown that engineered VSV with cell death elements has considerable antitumor efficacy. This chapter provides a comprehensive overview of VSV engineering techniques, focusing specifically on the incorporation of the proapoptotic gene XAF1, along with detailed experimental procedures. The goal of these approaches is to increase the efficacy of VSV in antitumor immunity and other therapeutic domains.