{"title":"Protocol for investigating the impact of transcription regulator deficiency on tumor-specific CD8<sup>+</sup> T cell responses via adoptive cell transfer.","authors":"Luming Xu, Yuzhu Wang, Mengqu Ran, Wen Liu, Lilin Ye, Qizhao Huang","doi":"10.1016/j.xpro.2025.103703","DOIUrl":null,"url":null,"abstract":"<p><p>Transcription factors play a crucial role in the differentiation of tumor-specific CD8<sup>+</sup> 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<sup>+</sup> T cells, induced by tamoxifen. We describe steps for examining the impact of ID3 deficiency on the differentiation of tumor-specific memory CD8<sup>+</sup> T cells (Ttsm) and progenitors of exhausted CD8<sup>+</sup> 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.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 2","pages":"103703"},"PeriodicalIF":1.3000,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"STAR Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.xpro.2025.103703","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
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.