Genome-wide CRISPR screening identifies the E3 ubiquitin ligase HERC1 as a modulator of the response to nucleoside analogs in acute myeloid leukemia. Genome-wide CRISPR screening identifies the E3 ubiquitin ligase HERC1 as a modulator of the response to nucleoside analogs in acute myeloid leukemia

We identified the Ubiquitin Ligase HERC1 in the context of cytarabine response in a genome-wide CRISPR screen in murine AML. We hypothesized to detect early transciptomic changes in cytarabine treated murine AML cells genetically targeted by using single guide RNAs targeting HERC1 and Non-targeting Control. Overall design: Early Detection of transcriptomic changes in cytarabine treated (200nm) murine MLL-AF9 transformed progenitor cells which were genetically modified with single guide RNA targeting HERC1 or Non-Targeting Ctrl RNA. AML clones targeted for HERC1 were sorted by using BD FACSAria II, and clone E5 was chosen for RNA sequencing. sgHERC1 - and sgCtrl MLL/AF9 cells were treated for six hours with 200nM cytarabine or H20 and RNA was subsequently isolated by using TRIzol (Invitrogen) + RNeasy mini kit (Qiagen, #74106).