Total RNA from edited and unedited CAR T cells was extracted after three days post αCD3-αCD28 activation with the High Pure RNA Isolation Kit (Roche) according to the manufacturer's recommendation and later purified and concentrated with the RNA Cleanup Kit (Monarch). The RNA quantity and purity (A260/ A280) ratio were measured on a Nanodrop spectrophotometer (Thermo Fisher Scientific), and the RNA integrity number (RIN) was determined using a 2100 Bioanalyzer with an Agilent RNA 6000 Pico kit (Agilent). mRNA-Sequencing was carried out by Novogene (UK) Company Ltd. Sample quality control, including RIN, was assessed with an Agilent 5400.  Subsequently , strand-specific library preparation, including polyA-enrichment, was performed according to their standard protocols and 30M paired-end reads were sequenced on an Illumina NovaSeq X Plus sequencer, using paired-end 150-bp sequencing strategy.