In vivo CRISPR screening in autoimmune disease model reveals differential dependence of Th1 and Th17 cells on amino acid transporter SNAT1

Purpose: The aim of this experiment was to determine the effect of SNAT1 loss on Th1 and Th17 cells. Methods: SNAT1 was knocked out using CRISPR/Cas9 in primary CD4+ T cells polarized to Th1 and Th17 cells in vitro. RNA was then isolated for RNAseq. n=3 biological replicates. Results: SNAT1 mRNA expression was significantly reduced in both Th1 and Th17 cells. Th1 cells displayed consequent changes in the mRNA expression of many other genes (with particular decrease in the expression of glycolysis pathways genes), while Th17 cells had no significant changes in the expression of all other mRNAs detected. Conclusion: This study demonstrates the differential dependence of Th1 and Th17 cells on the amino acid transporter SNAT1. mRNA profiles of Th1 and Th17 cells with non-targeting control (NTC) or SNAT1 gRNA