Establishment of leukemia models by CRISPR/Cas9–mediated gene editing of HSPCs. (A) Frequency of mutations in human AEL¹⁰  of genes edited by CRISPR/Cas9. (B) Patterns of mutational cooccurrence and mutual exclusion in the 10 genes selected for gene editing. In orange are cell cycle regulators; in red, genes regulating DNA methylation; in purple, transcription factors; in pink, chromatin regulation genes; and in green, the cohesin gene STAG2. (C) Kaplan-Meier survival curves of established leukemia mouse models and control mice (nontargeting guide [NTG]). Mice from primary, secondary, tertiary, and quaternary serial passages are shown. Survival comparison was analyzed by log-rank test. (D) Spleen weight (left) from primary (P, n = 14), secondary (S, n = 42), tertiary (T, n = 15), and quaternary (Q, n = 16) recipient mice and from control mice (NTG). The mean weight is shown by the horizontal line in the scatter dot plot, and the error bars represent the standard deviation. (E) Representative histology showing multisystemic infiltration of representative leukemia models of AEL (#744), mixed AEL/B-ALL (#595), and T-ALL (#755). Necropsy revealed a more extensive meningeal leukemic infiltration of meninges in lymphoid models compared with erythroid models. Images are liver, kidney, and brain (meninges). Scale bars in column 2 are 50 μm; the remainder are 100 μm. (F) Changes in gRNA enrichment in HSPCs pretransplant and in developed tumors as determined by amplicon sequencing and deep targeted sequencing. (G) Induced CRISPR/Cas9 mutations in Trp53, Bcor, and Dnmt3a. Plotted data are from supplemental Table 11. ns, not significant.