Peripheral CD8+ T cell tolerance and exhaustion are immunoregulatory checkpoints that attenuate effector T cell responses in both autoimmunity and cancer. Although each state is molecularly distinct and arises from fundamentally different points of the immune response, some phenotypic features of peripheral tolerance and exhaustion overlap. This raises the possibility that similar molecular processes drive both states, which has implications for how they might be targeted by immunotherapies in the clinic. Notably, the transcription factor, TOX, a master regulator of exhaustion, is also up-regulated in tolerant cells, suggesting that a core shared molecular and epigenetic program underlies both processes. We thus sought to both validate that TOX is induced during peripheral tolerance, and to probe its role in the tolerance process. CD8+ T cells undergoing peripheral tolerance induction expressed higher levels of TOX protein compared to early effector CD8+ T cells responding to infection, however TOX expression in tolerant cells was markedly lower than in exhausted CD8+ T cells. To dissect whether TOX functionally enforces tolerance, we knocked out TOX within CD8+ T cells by CRISPR editing and assessed its role within in vivo peripheral tolerance models. Strikingly, deletion of TOX in tolerant cells did not impair tolerance induction nor promote effector differentiation, even in the context of bystander inflammation and elevated antigen load. Thus, TOX is dispensable for peripheral tolerance, highlighting a fundamental molecular difference between the transcriptional and epigenetic programs driving tolerance and exhaustion. This provides functional proof that tolerance and exhaustion are molecularly distinct processes, which has broader implications in the design of next-generation immunotherapies.