Ets1 dampens expression of Tbet and CD8- or NK-like effector programing in postselection iNKT1 cells [scRNA-seq]

The goal of this study was to compare the gene expression program of thymic iNKT1 cells from Control mice to those with a postselection deletion of the transcription factor Ets1 at the single cell level. Ets1-floxed alleles were deleted in iNKT1 cells using Tbx21Cre bacterial artifical chromosome transgenic mice that also carry a Rosa26-floxed-stop-YFP reporter. Control iNKT1 cells were Tbx21Cre+ Rosa26-floxed-stop-YFP reporter+. We isolated all thymic YFP+ cells from Control and cKO mice and processed for scRNA-seq using the 10X Genomics Chromium. Data was processed using the Cell Ranger program for 10X Genomics. . We found that iNKT1 cells lacking Ets1 upregulated Tbx21 and its protein Tbet, many Tbet target genes, and CD8 effector T cell associated genes. Expression profiling analysis of RNA from thymic iNKT1 cells isolated from Ctrl (Tbx21Cre; Rosa26-YFP+) and cKO (Tbx21Cre; Rosa26-YFP+; Ets1F/F) mice by single cell RNA-sequencing.

本研究旨在在单细胞水平上，对比对照组小鼠与转录后阶段特异性缺失转录因子Ets1（Ets1）的小鼠的胸腺iNKT1细胞（thymic iNKT1 cells）的基因表达程序。研究人员通过同时携带Rosa26-floxed-stop-YFP报告基因（Rosa26-floxed-stop-YFP reporter）的Tbx21Cre细菌人工染色体转基因小鼠，在iNKT1细胞中敲除Ets1-floxed等位基因。对照组iNKT1细胞的基因型为Tbx21Cre+ Rosa26-floxed-stop-YFP reporter+。研究人员从对照组与条件性敲除（cKO）小鼠中分离所有胸腺YFP阳性细胞，并依托10X Genomics Chromium平台开展单细胞RNA测序（single cell RNA-sequencing，简称scRNA-seq），测序数据采用10X Genomics配套的Cell Ranger软件进行处理。研究发现，缺失Ets1的iNKT1细胞会上调Tbx21及其编码蛋白Tbet、大量Tbet靶基因以及CD8效应T细胞相关基因的表达水平。本研究通过单细胞RNA测序技术，对取自对照组（Tbx21Cre; Rosa26-YFP+）与条件性敲除组（Tbx21Cre; Rosa26-YFP+; Ets1F/F）小鼠的胸腺iNKT1细胞的RNA进行了表达谱分析。