Despite the success of CAR T cell-based immunotherapy, a sizable proportion of patients treated with CAR T cells experience disease relapse.  This may be due to the low persistence or exhaustion of CAR T cells resulting from their continuous exposure to the tumor antigen. Several genes have been implicated in pathways that control T cell exhaustion, acting either as promoters or as antagonists of this mechanism. In an effort to generate CAR T cells that resist T cell exhaustion, we sought to establish a robust genome editing procedure for multiplexed editing of CD19-specific CAR T cells. First, we identified the optimal conditions for manufacturing large quantities of CD19-specific CAR T cells using lentiviral-mediated transgene delivery. We tested several protocols and identified the conditions that consistently yielded over 50% of CAR-positive T cells. We then established an assay to induce CAR T cell exhaustion in vitro through tonic CAR signaling, and used this system to identify genes that either promote or antagonize exhaustion using transcriptome analyses of CAR T cells collected at different time points. A variety of genome editing strategies are currently being tested to achieve our ultimate goal of manufacturing CAR T cells with reduced expression of genes that promote exhaustion while simultaneously increasing the expression of genes that prevent exhaustion.  This study opens novel opportunities towards the generation of improved allogeneic cancer immunotherapies and might potentially extend the therapeutic use of this platform to more resilient tumor types in the future.