Fig. 1

Genome-wide CRISPR/Cas9 screening identifies RNA splicing factor RBM5 as a novel regulator for HOXA9 expression in acute leukemia. a Schematic diagram of a working flow of loss-of-function CRISPR screening targeting whole genome gene. b The whole-genome-wide CRISPR/Cas9 screen results. Bubble plots show the 19,063 genes identified, genes are known to regulate HOXA9 (HOXA9, HOXA7, ZFP64, USF2), and the novel hits RBM5, TADA3, SFSWAP, and CLASRP are highlighted in red. The P-values were calculated using the RSA algorithm. c The count ratio for all sgRNAs targeting the HOXA7, USF2, ZFP64, RBM5, TADA3, SFSWAP, and CLASRP is shown between HOXA9^High and HOXA9^Low sorted populations. The DEseq2 score of each gene was calculated by Log₂[fold change (HOXA9^High/HOXA9^Low)]. d Gene Ontology (GO) analysis was performed on the significant genes that positively regulate HOXA9 expression from the screening results. e RBM5 mRNA expression level in various cancer cell types from CCLE (Cancer Cell Line Encyclopedia). TPM, transcripts per million. T, tumor/cancer. f Box plot comparing the RBM5 mRNA expression level between AML samples (from TCGA dataset) and matched normal samples (from TCGA and GTEx projects). The plot is drawn by using the GEPIA online server. TPM, transcripts per million. T, tumor/cancer. N, normal bone marrow. *P < 0.05, unpaired Student’s t-test