The KDM2B Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population disrupting the KDM2B gene in the near-haploid HAP1 cell line. This pooled population comprises a heterogeneous mix of edited cells, offering a robust loss-of-function model that avoids the biases and artifacts of clonal selection. Without requiring single-cell cloning, these cells facilitate direct phenotypic and molecular analyses, making them immediately suitable for functional genomics and high-throughput epigenetic screens.
HAP1 is a male-derived, near-haploid human cell line originating from the KBM-7 chronic myeloid leukemia (CML) line. It maintains a haploid karyotype for all chromosomes except a duplication of chromosome 8 and displays a fibroblast-like morphology. Due to its genetic simplicity and stable growth, HAP1 is a preferred host for CRISPR-based functional genomics, facilitating clear genotype-phenotype correlations and scalable assay conditions. Its leukemic provenance provides a disease-relevant context for cancer studies.
KDM2B encodes a histone demethylase that specifically removes H3K4me3 and H3K36me2 marks at CpG islands, acting through its JmjC domain. It recruits a non-canonical Polycomb repressive complex 1 (PRC1) containing RING1B, PCGF1, and BCOR to catalyze histone H2A monoubiquitination, silencing target genes such as HOX clusters and the CDKN2A tumor suppressor. Additionally, KDM2B functions as an F-box protein within an SCF ubiquitin ligase complex composed of SKP1 and CUL1, suggesting connections to protein turnover and cell cycle regulation.
In the HAP1 CML background, KDM2B knockout enables precise dissection of oncogenic epigenetic reprogramming. KDM2B is frequently overexpressed in leukemia, lymphoma, and solid tumors, driving proliferation and blocking differentiation. Its loss triggers senescence, cell cycle arrest, and apoptosis. The near-haploid nature of HAP1 amplifies such phenotypic effects, facilitating sensitive detection of changes in viability and gene expression, and establishing a robust platform for investigating KDM2B-dependent tumorigenic mechanisms.
These polyclonal knockout cells are validated for a range of assays: western blotting for KDM2B and histone modifications (H3K4me3, H3K36me2, H2AK119ub); ChIP-qPCR at CpG island promoters to assess PRC1 occupancy; and RT-qPCR for expression changes in HOX genes and CDKN2A. Proliferation, apoptosis, and cell cycle assays further support drug target validation and functional epigenomic screens. This product serves as a versatile tool for epigenetic regulation studies and cancer biology. For inquiries, contact Ascent Research.
