The KDM6A Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered for targeted disruption of the KDM6A gene. This heterogeneous pool of HEK293T cells carries CRISPR/Cas9-mediated gene edits, providing a loss-of-function model without clonal selection. KDM6A encodes a histone demethylase that removes methyl groups from H3K27me3/me2, facilitating gene activation, and its knockout enables investigation of epigenetic regulation and transcriptional control.
HEK293T cells are an adherent human embryonic kidney derivative constitutively expressing the SV40 large T-antigen, which ensures high transfection efficiency and robust viral production. This line is widely used for recombinant protein expression and functional genomics. The cellular background retains active Notch, retinoic acid, and TGF-beta signaling cascades, making it relevant for studying KDM6A-mediated transcriptional regulation in a near-physiological signaling context.
KDM6A functions as a transcriptional coactivator by demethylating H3K27me3/me2, thereby relieving gene silencing. It cooperates with MLL3 (KMT2C) and MLL4 (KMT2D) methyltransferase complexes that catalyze H3K4me3 deposition. Upstream regulators include retinoic acid receptors (RAR/RXR), Notch intracellular domain (NICD), SMAD2/3, and p53. KDM6A transcriptionally activates downstream targets such as HOX gene clusters, CDKN1A (p21), BMP4, and GATA6. It physically associates with the SWI/SNF complex (via BRG1/BRM) and MLL3/4 core components ASH2L, RBBP5, WDR5, and DPY30.
In HEK293T cells, KDM6A disruption provides a model to dissect its tumor-suppressive and developmental roles. The knockout recapitulates loss-of-function conditions relevant to Kabuki syndrome, acute myeloid leukemia, bladder cancer, colorectal cancer, and breast cancer. Because these cells harbor functional p53 and TGF-beta pathways, researchers can examine how KDM6A integrates signals from NICD, SMAD2/3, and p53 to modulate gene expression. Analysis of H3K27me3 dynamics at target loci becomes straightforward in this system.
This product is suited for epigenetic studies, drug target validation, and disease modeling. Representative assays include Western blotting for knockout confirmation, ChIP-qPCR for H3K27me3 enrichment at promoters, RNA-seq for transcriptome profiling, and reporter gene assays for pathway activity. Cell proliferation and viability assays can assess functional consequences, and high-throughput screening applications are supported. For detailed technical support or customized protocols, please contact Ascent Research.
