is frequently mutated in acute myeloid leukemia (AML). To explore the features of human AML with the hotspot R882H mutation, we generated Dnmt3a R878H conditional knockin mice, which developed AML with enlarged LinSca1cKit cell compartments. The transcriptome and DNA methylation profiling of bulk leukemic cells and the single-cell RNA sequencing of leukemic stem/progenitor cells revealed significant changes in gene expression and epigenetic regulatory patterns that cause differentiation arrest and growth advantage. Consistent with leukemic cell accumulation in G/M phase, CDK1 was up-regulated due to activation associated with DNA hypomethylation. Overexpressed CDK1-mediated EZH2 phosphorylation resulted in an abnormal trimethylation of H3K27 profile. The mTOR inhibitor rapamycin elicited a significant therapeutic response in Dnmt3a mice.