Fig. 5
From: Development of drug-inducible CRISPR-Cas9 systems for large-scale functional screening
FACS-based pooled CRISPR screens using drug-inducible sgRNA expression constructs. (a) Schematic overview of FACS-based CRISPR screening approach to identify PD-L1 expression regulators. (b) Regulation of PD-L1 expression by IFNγ signaling pathway. Known positive PD-L1 regulators including PD-L1 itself are shown in red. Known negative PD-L1 regulators are shown in blue. (c) Scatter plot for the result of constitutive CRISPR screen. Each dot indicates median log2 fold change of all sgRNAs for one target gene. (d) Scatter plots for the screening results using DOX- or IPTG-inducible sgRNA vectors. Each dot indicates median log2 fold change of all sgRNAs for one target gene. 1 μg/mL DOX or 1 mM IPTG was used to induce the sgRNA expression. (e,f) Frequency histograms of the changes of sgRNA abundance in PD-L1low (e) and PD-L1high (f) cells versus pre-sort. sgRNAs targeting known PD-L1 positive regulating genes are shown by red and sgRNAs targeting known PD-L1 negative regulating genes are shown by blue