Modeling_Asxl1mut_Clonal_Hematopoiesis_using_32D_cell_line_

ASXL1 mutations characterize a subgroup of adverse-risk acute myeloid leukemia (AML) and are also common in other types of myeloid malignancies (MM). To identify therapeutic vulnerabilities of ASXL1-mutant MM, we generated ASXL1mut isogenic cell lines using CRISPR-Cas9 genome editing starting with murine myeloblast-like 32D cells. Using gRNAs against murine Asxl1 Exon 13, we generated pathogenic frameshift/stop mutations (recapitulating what is seen in human MM) and isolated several single cell-derived mutant and control clones for downstream applications. As part of this, we conducted a genome-wide CRISPR-Cas9 drop-out screen on mutant vs control 32D cell lines (clones) to identify genetic vulnerabilities specifically associated with ASXL1mut. Potential hits will be validated using in-vitro models and primary samples to evaluate their potential as anti-AML therapeutic targets.