Fig. 1: CRISPR screen of cPRC1-dependent gene silencing reveals Pds5a.
From: Loss of cohesin regulator PDS5A reveals repressive role of Polycomb loops
a Schematic of ectopic dual reporter locus consisting of 7x TetO landing sites flanked by an upstream Ef1a promoter driven BFP and a downstream PGK driven puromycin/GFP (top). Genomic ChIP-CapSeq screenshot of PcG proteins and histone modifications before (black) and after ectopic TetR-Cbx7 expression (orange) (bottom). Also see Supplementary Fig. 1a and Supplementary Table 3. b Flow cytometry histograms of GFP signal before (left) and after 4 days of Dox-dependent reversal of TetR-CBX7 tethering (right) in control (top) and Ring1b KO (bottom) Polycomb reporter ESCs. Percentages refer to fraction of GFP-negative ESCs. c Schematic of CRISPR screen design. MOI = multiplicity of infection. Genes (sgRNA library described in ref. 32) are rank-ordered based on CRISPR significance score (−log 10 MAGeCK significance score32; n = mean of three independent experiments). d Flow cytometry histograms of GFP signal before (left) and after 4 days of Dox-dependent reversal of TetR-CBX7 tethering (right) in control (top) and Pds5a KO (bottom) Polycomb reporter ESCs. Percentages refer to fraction of GFP-negative ESCs.
