Fig. 3: Cas9-mediated disruption of the h-FLT3 enhancer blunted MLL-r leukemia cell growth.

ChIP-seq tracks of transcription factors and H3K27ac (GEO: GSE117864)⁷⁰ were used to demonstrate the epigenetic status of the FLT3 promoter (A) and the HOXA9-bound site in the h-FLT3 enhancer (B). C Two sgRNAs (sgFLT3-DE-1 and -DE-2) targeting the h-FLT3 enhancer were used to disrupt the h-FLT3 enhancer activity. D Q-PCR detection of gene expression demonstrated notable downregulation of FLT3 and PAN3 expressions in SEM cells upon CRISPR targeting. Cas9/sgFLT3-DE-1/2-mediated disruption of the h-FLT3 enhancer resulted in retarded cell growth of MLL-r leukemia cells (E–G) but not non-MLL-r leukemia cells or OCI-AML-2 cells with low enhancer activity of the h-FLT3 (H–J). The sgRPS19-11 sgRNA served as the positive control. The percentage of cell numbers was normalized to CFP⁺ control cells (sgNT). Data are shown as mean values ± SEM of three biological replicates (center of the error bar). P values were estimated using a two-tail un-paired t-test. Source data are provided as a Source Data File.