CRISPR-Cas9 sgRNA screening for DUB family genes regulating AE fusion protein in t(8;21) AML cells

This project investigates the role of deubiquitinases (DUBs) in regulating the stability of the AE fusion protein in t(8;21) acute myeloid leukemia (AML). A custom CRISPR-Cas9 sgRNA library was designed containing 960 sgRNAs targeting 96 DUB family genes and delivered into Skno-1 cells via lentiviral transduction. Following puromycin selection, cells were clonally expanded and subjected to next-generation sequencing (NGS) and protein analysis. Genomic sequencing was performed to identify DUB genes associated with AE degradation. The results highlight USP5 as a critical regulator positively correlated with AE expression in AML cells. Raw sequencing data generated from this screen are deposited in the Sequence Read Archive (SRA).