Fig. 5: Identification and characterization of hTSC GRGs.

a Growth-restricting genes (GRGs) identified among all targeted genes. b The mean of sgRNA normalized read counts and Log2 fold changes of all or selected hTSC GRGs and their neighboring up- and downstream genes over time. The results are representative of two independently transduced screening experiments. c Top 5 most enriched GO Biological Processes terms among all hTSC GRGs. d Top left, live cell counts relative to the mean of control sgRNA transduced H9 hTSCs 6 days after seeding. Error bar represents the standard error of three biological replicates. Two-tailed student’s t test was used for statistical analysis. “*” indicates a p-value < 0.05, “**” indicates a p-value < 0.01, and “***” indicates a p-value < 0.001. The exact p-values from left to right are 0.000870208, 0.005413584, 0.018757677, and 0.000646371. Rest of the panel, phase contrast images of H9 naïve hTSCs transduced with control or targeting sgRNAs at 6 days following seeding. The scale bars indicate 75 μm. The images are representative of three independent replicates in an experiment. e Heatmap showing the expression of 50 CTB-enriched GRGs in the CTB, EPI, and PrE of published 10 d.p.f. human embryo scRNA-seq data³⁷. f Dot plot showing the expression of selected CTB-enriched GRGs in the CTB, EPI, and PrE of published human embryo scRNA-seq data³⁷. g Heatmap showing the expression of 33 CTB-depleted GRGs in the CTB, EPI, and PrE of published 10 d.p.f. human embryo scRNA-seq data³⁷. h Dot plot showing the expression of selected CTB-depleted GRGs in the CTB, EPI, and PrE of published human embryo scRNA-seq data³⁷.