Fig. 1: DDR1 was identified as an ICI resistance gene by in vivo CRISPR screening.

a CRISPR sgRNA library screening flow chart. b Round 1: We identified downregulated sgRNAs in pool A that could sensitize the PD-1 blockade effect by the CRISPR/Cas9 method (n = 5-6/each group). c Round 2: Mouse model experiments were conducted to verify the top 8 candidate genes in pool A (n = 5-6/each group). d Tumor inhibition rate of tumor volume in Round 2 with knock out effect of 8 candidate genes on CT26 mouse models. e RNA-sequencing analysis results of pool A.Heatmap (left) showing the downregulated kinase genes with their specific sgRNAs in sample 1 and sample 2 in the PD-1 blockade group and control group. Venn diagram (right) showing the overlapping and top 8 ranked kinases downregulated in both sample 1 and sample 2. (p < 0.05;** p < 0.01;***, p < 0.001, n.s., not significant). f Kinome tree presenting protein kinases that were significantly downregulated in sample 1 and sample 2 (Pool A PD-1 blockade group). Dark blue circles represent the overlapping kinase genes that were downregulated in both sample 1 and sample 2 with Pool A sgRNA deleted and treated with PD-1 blockade (same as the following). The gray blue and light blue circles represent the kinase genes downregulated in sample 1 and sample 2, respectively. The red rectangle represents the kinase genes that were upregulated in both sample 1 and sample 2. The pink and purple rectangles represent the kinase genes upregulated in sample 1 and sample 2, respectively. Image created using KinMap, illustration reproduced courtesy of Cell Signaling Technology, Inc. (https://www.cellsignal.com).