SLAM-seq analysis of mRNA half-lives in human CD8? T cells during activation
SLAM-seq (thiol(SH)-linked alkylation for the metabolic sequencing of RNA) was applied to human primary CD8? T cells subjected to in vitro activation with aCD3/aCD28 beads. To quantify mRNA half-lives and their dynamic changes during CD8? T cell activation, we implemented a pulse-chase design with 24 hours of labelling using 100 µM 4-thiouridine (4SU), replenished every 4 hours with fresh RPMI medium containing 100 µM 4SU. This was followed by uridine washout for 0, 0.5, 1, 3, 6, and 16 hours. Samples were collected from non-activated, Day1-activated, and Day5-activated CD8? T cells at each washout step (2 donors per time point). These datasets provide transcriptome-wide measurements of mRNA half-lives and reveal how mRNA turnover alters upon T cell activation.