The KDM2A Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the KDM2A gene. This product provides a heterogeneous pool of HAP1 cells carrying targeted disruptions of KDM2A, generated via CRISPR/Cas9-mediated gene disruption without clonal isolation. The polyclonal format ensures a diverse representation of editing events, making it suitable for pooled functional assays and studies where clonal variability is not desired. By eliminating KDM2A expression, this model enables researchers to interrogate the gene??s role in epigenetic regulation, signal transduction, and cancer biology, offering a robust tool for mechanistic and screening applications.
The host HAP1 cell line is a near-haploid human cell line derived from the KBM-7 chronic myelogenous leukemia line, widely employed in functional genomics and genetic screens due to its simplified single-copy genome. Its haploid state facilitates straightforward knockout generation and genotype-phenotype correlation, minimizing complications from heterozygous mutations. HAP1 cells retain key signaling pathways and express many native proteins, making them a physiologically relevant model for studying gene function. The combination of this well-characterized host with CRISPR/Cas9-mediated KDM2A disruption provides a powerful system to investigate epigenetic mechanisms in a clean genetic background.
KDM2A encodes a histone demethylase that specifically removes mono- and dimethyl marks from lysine 36 of histone H3 (H3K36me1/2), primarily at promoter regions, leading to transcriptional repression. It forms repressive complexes with SIN3A, HDAC1, HDAC2, CoREST, and BHC80, linking demethylation to chromatin compaction. KDM2A is activated by upstream regulators such as the Notch intracellular domain (NICD), NF-??B (p65/RELA), p53, and hypoxia-inducible factor 1-alpha (HIF1A), and it represses key downstream targets including CDKN1A (p21), CCND1 (Cyclin D1), HES1, IL8, and CXCL2. Through these interactions, KDM2A integrates signals from Notch and NF-??B pathways to control cell cycle progression, inflammatory responses, and senescence.
Disruption of KDM2A in HAP1 cells abolishes its demethylase activity, leading to elevated H3K36me2 levels at genomic targets and derepression of silenced genes. This alters Notch and NF-??B target gene expression, impacting processes such as cell cycle arrest, senescence, and cytokine production. The near-haploid nature of HAP1 ensures that the knockout phenotype is directly linked to KDM2A loss without compensatory alleles, providing a clear readout of its epigenetic regulatory role. Consequently, this model is well-suited for dissecting how KDM2A-mediated histone modifications govern cellular responses to developmental and oncogenic signals.
Researchers can utilize these polyclonal knockout cells in a variety of experimental contexts, including functional genomics, epigenetic regulation studies, and cancer cell biology. Representative assays include ChIP-qPCR to assess H3K36me2 enrichment, western blotting for KDM2A protein levels, RT-qPCR for target gene expression, MTT proliferation assays, and luciferase reporter assays for Notch activity. The cells are also compatible with RNA-seq for transcriptome-wide analysis and immunofluorescence for localization studies. Applications extend to drug target screening and investigations of acute myeloid leukemia, breast cancer, and glioblastoma. For further technical details or ordering information, please contact Ascent Research.
