The KDM6A Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population specifically designed to disrupt the KDM6A gene in the HAP1 haploid human cell line. This pooled knockout model provides a genetically heterogeneous loss-of-function system for studying the epigenetic functions of KDM6A without clonal selection biases. The polyclonal format is well-suited for bulk population-based assays and high-throughput screening applications where functional diversity may reveal consistent phenotype trends. Researchers can rely on this knockout product as a versatile tool to investigate the consequences of KDM6A deficiency in a genetically tractable and reproducible cellular background.
HAP1 cells are a near-haploid fibroblast-like cell line derived from the KBM-7 chronic myeloid leukemia cell line. Their haploid karyotype simplifies the generation of knockout alleles by CRISPR/Cas9, as only one allele needs to be disrupted to produce a complete gene loss in the majority of cells. This characteristic makes HAP1 an ideal model system for genetic knockout studies, functional genomics, and large-scale screening endeavors. The retention of key signaling pathways and a stable mesenchymal morphology further support their application in dissecting gene regulatory networks relevant to cancer biology and cellular differentiation.
KDM6A encodes a histone H3 lysine 27 demethylase that removes di- and tri-methyl marks from H3K27me2/me3, relieving transcriptional repression. Within COMPASS-like complexes, it interacts with MLL3/4, ASXL1, and BAP1. Activated by retinoic acid and NICD, KDM6A demethylates H3K27me3 at promoters of downstream targets including HOXA cluster genes, CDKN1A (p21), NOTCH1, and RB1. This places KDM6A at a convergence point of Notch, TGF-beta, and retinoic acid signaling, where it facilitates expression of differentiation and tumor suppressor programs.
KDM6A knockout in HAP1 cells is predicted to cause accumulation of H3K27me3 at target gene promoters, leading to sustained repression of key loci. This alteration disrupts normal cell differentiation programs and may mimic aspects of Kabuki syndrome and certain cancers where KDM6A is frequently mutated. The near-haploid background allows clear detection of epigenetic changes and gene expression shifts without the confounding effects of a second wild-type allele. Consequently, this polyclonal knockout population serves as a powerful platform for mapping KDM6A-dependent transcriptomes and assessing the functional relevance of KDM6A-associated protein interactions in a simplified genomic context.
Applications include histone modification profiling by Western blotting or mass spectrometry to measure H3K27me3, ChIP?CqPCR to examine H3K27me3 at HOX gene promoters, and RT?qPCR or RNA?seq to quantify expression of downstream targets like CDKN1A and NOTCH1. The cells support proliferation and colony formation assays to assess growth effects, and drug sensitivity screening to identify synthetic lethal partners. The polyclonal format is ideal for high?throughput genetic and chemical screens, enabling systematic mapping of KDM6A?dependent vulnerabilities. For technical inquiries, please contact Ascent Research.
