Numerous types of cancer have been associated with the Epstein-Barr Virus (EBV), a herpesvirus that has infected more than 95% of the global population. Moreover, all identified EBV-related malignancies contain the Epstein-Barr nuclear antigen 1 (EBNA1). EBNA1 is a crucial viral protein that binds to host cell DNA to maintain itself and is necessary for viral replication and cell immortalization. Lacking known enzymatic functions, EBNA1 relies on interactions with other host cell proteins, such as ubiquitin-specific protease 7 (USP7). USP7 is a deubiquitinating enzyme that safeguards proteins from degradation via the ubiquitin-proteasome pathway; however, it remains uncertain whether USP7 is a proviral factor for EBV. Our hypothesis posits that inhibiting USP7 activity will negatively affect the growth of EBV+ cancer cells. We found that EBV+ cells treated with pharmacological inhibition of USP7 via GNE6776, a small molecule that blocks the catalytic cysteine of USP7, had significantly reduced levels of EBNA1 as measured by Western Blot. The pharmacological phenotype was replicated using alternative USP7 inhibitors– XL177A and (R)-FT671. Further analysis revealed that viral DNA was significantly decreased in EBV+ cells treated with USP7 inhibitors. To better understand how the reduction in EBNA1 and viral DNA could impact EBV+ cells, we analyzed how USP7 impacts cellular metabolism via resazurin assay. Our results showed that disrupting USP7 through various independent inhibitors selectively restricts the metabolic function of EBV+ cancer cells. Next, we examined how USP7 inhibition could impact cell division by performing cell cycle analysis. EBV+ cells treated with GNE6776 contained more cells in G1 phase and less in S phase, suggesting that USP7 inhibition also hinders cell cycle progression. While inhibiting USP7 alone could reduce cell growth, we tested whether USP7 inhibitors could potentiate the impact of an EBNA1 inhibitor that our lab has previously developed. To evaluate this hypothesis, we tested two novel bifunctional molecules, ARK-02-19 and ARK-02-58, synthesized from a USP7 inhibitor conjugated with an EBNA1 inhibitor. Resazurin assays following ARK-02-19 and ARK-02-58 treatment demonstrated preferential targeting of EBV+ cancer cells. In conclusion, pharmacological inhibition of USP7 effectively targets EBV+ cancer cells by reducing EBNA1 levels, thereby disrupting viral DNA replication. Additionally, we found that EBV+ cancer cells' growth is more sensitive to USP7 inhibitors than EBV- cancer cells. Our work demonstrates that disrupting USP7 is a promising, novel therapeutic strategy for EBV-associated cancers. Future work includes further evaluation of these novel bifunctional molecules and in vivo tests of USP7 inhibition via an orthotopic mouse model currently in development.