Effect of Grk2 (spleen, rat) or Ets1 (spleen and spinal cord parenchyma, rat; in vitro, human) KOs on gene expression of CD4+ T cells

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To study the effect of the Grk2 or Ets1 KOs in rat and human CD4+ T cells, in vivo or in vitro respectively, we generated KO CD4+ T cell lines by CRISPR-RNP nucleofection (with an gRNA targeting the gene locus) or control (with a non-targeting gRNA)

3' bulk RNAseq was conducted on CD4+ T-MBP rat cells isolated form spleen or spinal cord parenchyma three days after co-transfer of control and KO cells into the animal. Cells were FACS sorted based on their fluorescent protein reporters, BFP for control, and GFP for KOs. For human experiments, CD4+ T cells were isolated from buffy coats, KOed and restimulated in vitro, and 3' bulk RNAseq was performed. For rats, three biological replicates were used from three different animals, for humans, four biological replicates were used from four different donors