Fig. 1: Genome-wide KO screens identify regulators of human neuroectoderm differentiation speed.
From: Genome-wide CRISPR screen identifies Menin and SUZ12 as regulators of human developmental timing
a, Schematic of the neuroectoderm differentiation. NE, neuroectoderm. Dual SMADi, inhibition of SMAD signalling by co-treatment of SB431542 and LDN193189. b, Temporal expression of PAX6 RNA from bulk RNA-seq experiment (left, n = 3 independent differentiations) and quantification of GFP expression in the PAX6::H2B-GFP cell line from flow cytometry analysis (right, n = 4 independent differentiations) during neuroectoderm differentiation. Data are the mean ± s.d. c, Representative histogram plots for live GFP expression in the PAX6::H2B-GFP cell line undergoing neuroectoderm differentiation. d0, day 0 of induction. d, Schematic of the whole-genome CRISPR screen. e, Representative flow cytometry gating strategy to isolate PAX6-GFPhigh and PAX6-GFPlow populations. f, Waterfall plot of the top 100 GSEA enriched pathways from the ranked gene list, ordered by Z-score of PAX6-GFPhigh versus PAX6-GFPlow comparison (top ranks correspond to highly enriched genes in PAX6-GFPhigh population). Chromatin-related and mitochondrial metabolism-related pathways are highlighted in coloured dots. g, Scatter plot of the screen hits (P-value < 0.05; Z-score > 1) highlighted in green dots. P-values were calculated from edgeR’s exact test.
