The ASXL1 Knouckout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ASXL1 gene, an epigenetic regulator frequently mutated in myeloid malignancies. This polyclonal pool of HAP1 cells facilitates robust and reproducible loss-of-function analyses without clonal isolation, enabling assessment of diverse editing outcomes. It is particularly suited for functional genomics and epigenetic drug screening applications.
The HAP1 host cell line is a near-haploid chronic myeloid leukemia-derived line with an adherent fibroblast-like morphology. Originally derived from KBM-7, these cells retain a near-haploid karyotype and express hematopoietic markers, offering an ideal genetic background for knockout models. The single-copy genome ensures that disruption of the ASXL1 allele leads to complete loss of protein function, enhancing phenotype detection in polyclonal populations.
ASXL1 functions as a scaffold protein within the BAP1 deubiquitinase complex and Polycomb repressive complex 2 (PRC2), interacting with BAP1, EZH2, SUZ12, and EED to modulate histone H2A deubiquitination and H3K27 trimethylation. It transcriptionally regulates downstream targets including HOXA5, HOXA9, HOXA10, MYC, and CDKN1A. Disruption of ASXL1, mediated by cytokine-induced signals and hematopoietic transcription factors, leads to derepression of HOXA cluster genes and altered H3K27me3 levels, placing ASXL1 at the nexus of chromatin remodeling and transcriptional control in hematopoietic stem cells.
In the HAP1 context, ASXL1 loss recapitulates myeloid leukemogenesis hallmarks through epigenetic dysregulation, leading to oncogenic HOXA gene expression and aberrant proliferation. This model is highly relevant for studying ASXL1-mutant myelodysplastic syndromes, acute myeloid leukemia, chronic myelomonocytic leukemia, and myeloproliferative neoplasms. The near-haploid karyotype amplifies phenotypic consequences, enabling clear dissection of ASXL1-dependent signaling and synthetic lethal interactions, making this knockout population a valuable tool for mechanistic studies and therapeutic testing.
This ASXL1 knockout cell population is widely applicable in advanced research, supporting functional genomics via haploid genetic screens to identify epigenetic regulators, and epigenetic drug screening, particularly with EZH2 inhibitors, using colony formation, proliferation, and drug sensitivity assays. Downstream analyses include Western blotting for ASXL1 and interacting proteins, RT-qPCR for HOXA genes, RNA-seq, and ChIP-qPCR for H3K27me3. It also enables studies of hematopoietic differentiation and cytokine-induced signaling. For further information, please contact Ascent Research.
