Extended Data Fig. 1: sgRNAs targeting EZH2 CXC ___domain and the drug-binding pockets are the most enriched in CRISPR-suppressor scanning.
From: Drug addiction unveils a repressive methylation ceiling in EZH2-mutant lymphoma
a, Scatter plots showing fitness scores (y-axis) in Karpas-422 under vehicle treatment at week 8. Fitness scores were calculated as the log2(fold-change) sgRNA enrichment under vehicle normalized to the mean of the negative control sgRNAs (n = 58). The PRC2-targeting sgRNAs (n = 650) are arrayed by their predicted cut sites in the coding sequences (x-axis). Data represent mean of n = 3 replicates. b, Structural view of the GSK343 binding site with EZH2 residues I109 and N699 highlighted in red. GSK343 analog is shown in gold. PDB: 5LS6. c, Structural view of the EED226 binding site with EED residue Y365 highlighted in red. EED226 analog is shown in gold. PDB: 5K0M. d, Bar plots of wild-type (gray) and EZH2 I109_M110insM mutant (red) allele frequencies from CRISPR-suppressor scanning under 5-week GSK343 (1 μM) or EED226 (1 μM) treatment. e, Same as in d. but for wild-type (gray) and EED Y365F and Y365F/M366V mutant (red) allele frequencies. f, Schematic showing genotypes and bar plots of allele frequencies for mutations that are observed at frequencies of >2% in the gDNA encoding EZH2 surrounding the CXCdel loop for 6-week GSK343 (1 μM) treatment after transduction with sgA596/D597 in Karpas-422. Allele frequencies under vehicle (gray) or GSK343 (red). (top) Schematic depicts the secondary structure of C-terminal CXC ___domain and cysteine residues that coordinate the zinc ions. g, Same as in f. but for sgD597 transduction in Karpas-422. h, Same as in f. but for sgD597/H598 transduction in Karpas-422. i, Same as in f. but for sgD597 transduction in Pfeiffer. j, The conservation scores of each amino acid within and surrounding the EZH2 CXCdel loop are shown in colors ranging from variable (blue) to conserved (red) as calculated by ConSurf-DB24. (Top) Schematic depicts the secondary structure of C-terminal CXC ___domain and cysteine residues that coordinate the zinc ions. k, Comparison of amino acids homologous to the EZH2 CXCdel loop in various model organisms. Data in a, d-i are represented by mean of n = 3.
