Genome-scale CRISPRi Perturb-seq on both unstimulated and stimulated Jurkat cells reveals regulators of human T-cell signaling

The goal of the experiment is to identify genes involved in T-cell receptor (TCR) signaling and T-cell activation in human T cells. Overall design: The Jurkat E6 cell line, engineered to express dCas9-KRAB and transduced with a genome-wide CRISPRi CROP-seq library, was isolated through Fluorescence-activated cell sorting (FACS) based on the percentage of GFP positivity. Subsequently, the isolated cells underwent analysis using single-cell RNA sequencing.