Extended Data Fig. 3: In vivo T cell CRISPR screen.
From: ST3GAL1 and βII-spectrin pathways control CAR T cell migration to target tumors
a. Volcano plot shows comparison of differentially expressed genes in naïve vs. in vitro activated CD8 T cells. Red denotes genes increased and blue denotes genes decreased in T cells. P values were adjusted using the Benjamini–Hochberg method. b. Heatmap of RNA-sequencing data from naïve vs. in vitro activated CD8 T cell samples (n = 3). Heatmap represents top 1,316 genes chosen from highest 0.25% p-value summary. c. Retroviral construct used for sgRNA delivery. d. A schematic of the experimental design showing timeline of T cell stimulation, transduction, enrichment, and flow analysis. T cells were purified by FACS sorting at day 7 and i.v injected into recipient tumor bearing mice. e. Scatter plot of the enrichment of candidates (n = 5 sgRNAs per gene) showing that sgRNAs targeting essential genes (left) were significantly reduced in the samples after two representative in vivo screens, but not the non-targeting guides (right). f. Genes with positive beta score (Tmigratory) and negative beta score (TSequestered) (>2 standard deviations from the mean (red)) are highlighted in yellow and blue, respectively.
