Protocol for investigating the impact of transcription regulator deficiency on tumor-specific CD8+ T cell responses via adoptive cell transfer
Year: 2025
Type: article
Abstract: 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 • Instructions for the establishment of the B16.GP melanoma mouse model • Steps for preparing Id3 −/− P14 and Id3 fl/fl P14 cells by the administration of tamoxifen • Guidance on investigating the effect of Id3 deficiency on CD8 + T cell differentiation Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.
Source: STAR Protocols
Language: English
Topic: T-cell and B-cell Immunology
Subfield: Immunology
Field: Immunology and Microbiology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,550
Funders: Natural Science Foundation of Guangdong Province, Natural Science Foundation of Chongqing, National Outstanding Youth Science Fund Project of National Natural Science Foundation of China, Key Technologies Research and Development Program, National Key Research and Development Program of China, Science Fund for Distinguished Young Scholars of Chongqing
OpenAlex ID: W4408864512