Extended Data Fig. 3: Rationale for CRISPR-ChIP after knockout of essential genes.
From: CRISPR–ChIP reveals selective regulation of H3K79me2 by Menin in MLL leukemia
a) sgRNA negative selection competition assay in K562 Cas9 cells transduced with control sgRNA or two independent sgRNAs against RNA Pol II (Pol II). Percentage of sgRNA positive cells remaining over time. Data represents the mean from n=2 experiments. b) FACS histogram of BFP expression in K562 Cas9 cells transduced with control sgRNA or two independent sgRNAs against RNA Pol II. Analysis performed at day 5 post infection. c) FACS scatter plot of forward scatter (FSC) vs side scatter (SSC) in K562 Cas9 cells transduced with control sgRNA or two independent sgRNAs against RNA Pol II. Analysis performed at day 5 post infection. d) Schematic of vector systems used for sgRNA expression and destabilised GFP (GFP-PEST) expression. e) FACS histogram of GFP expression in K562 Cas9 EF1a-GFP-PEST cells transduced with control sgRNA or two independent sgRNAs against RNA Pol II. Analysis performed at day 5 post infection. f) FACS scatter plot of GFP vs BFP expression in K562 Cas9 EF1a-GFP-PEST cells transduced with control sgRNA or two independent sgRNAs against the RNA Pol II. Analysis performed at day 5 post infection. g) Correlation plot of normalised Pol II levels (rpm) vs normalised H3K4me3 levels (rpm).
