Protocol for investigating the impact of transcription regulator deficiency on tumor-specific CD8+ T cell responses via adoptive cell transfer

Transcription factors play a crucial role in the differentiation of tumor-specific CD8⁺ T cells and significantly influence their capacity to repress tumors. Here, we present a protocol for creating a transcription regulator inhibitor of DNA 3 (ID3) conditional knockout mouse in tumor-specific CD8⁺ T cells, induced by tamoxifen. We describe steps for examining the impact of ID3 deficiency on the differentiation of tumor-specific memory CD8⁺ T cells (Ttsm) and progenitors of exhausted CD8⁺ T cells (Tpex) in tumor-draining lymph nodes through a co-adoptive transfer assay. For complete details on the use and execution of this protocol, please refer to Ran et al.¹.