DDX6 undergoes phase separation to modulate metabolic plasticity and drug resistance in AML

DDX6 is a member of the DEAD-box helicase and it is highly expressed in acute myeloid leukemia (AML). The Knockout (KO) of DDX6 in AML cells significantly suppressed their growth, reduced their mitochondrial oxidative phosphorylation (OXPHOS) in vitro, and substantially delayed leukemogenesis in vivo. To delineate the molecular mechanism through which DDX6 exerts its biological functions in AML, we conducted transcriptome-wide RNA sequencing with Mono-mac-6 AML cells following DDX6 KO. We introduced the single-guide RNAs (sgRNAs) targeting DDX6 (sgDDX6-1 and sgDDX6-2) or the control sgRNA (sgNS) into Cas9 single-clone of Mono-mac-6 AML cells via lentivirus transduction. The transduction-positive AML cells were selected with blasticidin (10 μg/ml; Cat #: ant-bl-1, InvivoGen) and hygromycin B (200 μg/ml; Cat #: ant-hg-5, InvivoGen) for 1-2 weeks. DDX6 KO efficacy was confirmed by Western blot. Then, the total RNAs were extracted from each group with QIAzol and the poly(A)+ RNAs were enriched from total RNAs for library construction and sequencing. For each group, we included 2 replicates.