FIG 2.

Genome-wide CRISPR/Cas screen. (A) Outline of screening strategy. (B) Flow cytometric analysis of J-dual#3 and J-dual#6 reporter cells without transduction and 4 days after transduction by the GeCKO v2 lentiviral vector library (MOI, around 2) immediately prior to the sorting process. The total number of events shown in the graph, total number of cells analyzed in the screen, and total numbers of cells sorted are indicated. (C) Enrichment analysis for guide RNAs in transduced and selected J-dual#3 and J-dual#6 reporter cells compared to transduced but unselected Jurkat cells by the MAGeCK program. The robust ranking aggregation (RRA) score reflects the enrichment of guide RNA targeting the same gene in cells selected from both reporter cell clones. The distribution of the negative log₁₀ of the RRA score for all targeted genes in alphabetical order on the x axis is shown, with the top 12 candidate genes highlighted. (D) Additional MAGeCK ranking parameters of the top 12 candidate genes. fdr, false discovery rate; # sgRNA, number of enriched guide RNAs targeting the same gene.