Bulk RNA sequencing of human FMC63 CAR T cells with and without overexpression of cystatin A (CSTA)
FMC63-based CAR T cells with or without CSTA overexpression from two independent productions were thawed and cultured in T-cell media + 40IU/ml IL-2 for 48 hours. After 48 hours, CAR T cells were washed with FACS buffer, stained with anti-CD3, anti-HA, and DAPI, and sorted for CAR expression. Sorted CAR T cells were then pelleted and flash frozen. RNA was extracted, polyA-enriched, and sequenced using Illumina NovaSeq S4 PE100 sequencing targeting 10 million read pairs per