The Cul5 E3 Ligase Complex Is a Key Negative Feedback Regulator of TCR/IL2 Signaling and Anti-Tumor Activity in CD8+ T Cells

To help advancing cancer immunotherapies, we developed a step-wise CRISPR knockout (KO) screening strategy under the selection of TGF-beta, and identified Cul5 as a core negative-feedback regulator of CD8+ T cell differentiation and persistence. Overall design: To comprehensively identify gene candidates regulating functional CD8+ T cell subgroups, we performed single-cell CRISPR KO screening with tumor-infiltrating CD8+ T cells by single-cell RNA sequencing in parallel with single-cell sgRNA sequencing.