Establishment of inducible CRISPR/Cas9 system in human AML cell lines

MOLM‐13 cell line was infected by pCW‐Cas9, doxycycline‐inducible lentiviral expressing Cas9 (Addgene plasmid #50661), and positively infected cells were selected by 2 μg/ml puromycin. sgRNAs (Table EV4) targeting p90RSK and CREB were cloned into pLKO5.sgRNA.EFS.tRFP657 (Addgene plasmid #57824). sgRNA lentivirus was prepared and used to infect inducible Cas9‐expressing human AML cell lines. Positive sgRNA‐infected cells were sorted based on tRFP657 expression. Cas9 was induced by 2 μg/ml doxycycline, and the cells were harvested 2 days after induction and subjected to subsequent analyses.