A custom CRISPR screen for genetic dependencies in CALM-AF10 AML. (A) Schematic of the CRISPR screen. A custom pooled library of sgRNAs targeting each of the 420 CALM-AF10 target genes and protein interactors was generated. Five sgRNAs targeting the early constitutive exon and 5 targeting functional domains were combined for a total of 10 sgRNAs per gene. sgRNA library virus was transduced in CALM-AF10-Cas9 AML cells and sgRNA representation was determined after in vitro culture or from AML cells obtained from terminally ill mice. (B-C) Plot showing the combined P values from 2 replicates (y-axis) of sgRNAs ranked in the order of depletion (x-axis) in the T₀ compared with the T₁₂ (B) or T_D (C) time points. Statistically significant hits are zoomed in and displayed in a separate graph on the right of each figure. JAK1, ranked ninth in vitro and sixth in vivo, is labeled in red. (D) Venn diagram of in vitro hits (yellow oval) and in vivo hits (red oval) are shown with overlapping genes displayed in the union of the 2 sets.