Massively parallel engineering of persistent CAR-Ts [CLASH]

To identify immune-related gene perturbations that enhance CAR-T cell persistence in in vitro and in vivo settings, we generated thousands of CAR-T variants using the CLASH platform and evaluated crRNA representation across time in in vitro and in vivo cancer models. After genomic DNA was collected from CAR-Ts (either from in vitro or in vivo experiments), a two step PCR was performed to generate a sequenceable guide library. Raw counts were processed using custom bash and R scripts.