circIkzf1 and Ikzf1 overexpression in Th cells [scRNA-seq B]

Cd4+ cells were isolated from mouse spleens, activated and retrovirally transduced to simultaneously express circIkzf1 and Ikzf1 or the respective controls to see how these two molecules would interfere with each other Overall design: Sequencing of RNA was performed from flow cytometry-sorted double-transduced living mouse Th cells (hCD4+ adn GFP+) expressing circIkzf1, Ikzf1 or controls, using the 10X Genomics platform with the Single Cell 5' Library & Gel Bead Kit. scRNA-seq sets A and B are technical replicates.

从小鼠脾脏中分离CD4+细胞，经活化后通过逆转录病毒转导，使其同时表达circIkzf1与Ikzf1，或相应的对照载体，以探究这两种分子间的相互调控作用。整体实验设计：采用10X Genomics平台及单细胞5'文库与磁珠试剂盒（Single Cell 5' Library & Gel Bead Kit），对经流式细胞术分选的双转导活小鼠辅助性T（Th）细胞——即同时表达hCD4+与GFP+，且分别转染circIkzf1、Ikzf1或对照载体的细胞——进行单细胞RNA测序（single-cell RNA sequencing，scRNA-seq）。其中scRNA-seq数据集A与B为技术重复。