The KDM6B Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HT29 colorectal adenocarcinoma line. This product features targeted disruption of the KDM6B gene, which encodes a histone H3K27me3 demethylase (JMJD3). The polyclonal format yields a heterogeneous loss-of-function model, enabling investigation of KDM6B-dependent epigenetic phenotypes without clonal selection bias.
The HT29 cell line is a widely used epithelial model of colorectal cancer, harboring oncogenic BRAF V600E, TP53, and APC mutations, and exhibiting microsatellite stability. These cells retain some capacity for enterocytic differentiation and endogenously express KDM6B, providing a disease-relevant background to study how H3K27 demethylation integrates with mutant signaling networks in colorectal tumorigenesis.
KDM6B specifically removes trimethyl marks from histone H3K27, antagonizing PRC2 (EZH2, SUZ12, EED)-mediated transcriptional repression. It is transcriptionally regulated by upstream factors including NF-??B (RELA), TGF-??/SMAD2/3, NOTCH intracellular domain, STAT3, retinoic acid receptor, p53, and TET proteins. The demethylase interacts with MLL3/4, SWI/SNF (BRG1), TET1, ASCL1, and SMAD3, and directly promotes expression of target genes such as the cyclin-dependent kinase inhibitors CDKN1A (p21) and CDKN2A (p16INK4a), HOXA cluster genes, BMP2, BMP4, NOTCH1, HES1, and VEGFA.
In the HT29 background, KDM6B knockout raises H3K27me3 levels at PRC2-bound loci, reinforcing silencing of differentiation and senescence programs. This effect, in conjunction with constitutive BRAF-MAPK signaling and p53 deficiency, may exacerbate the undifferentiated state and modulate WNT/??-catenin and NOTCH pathway activities. The model is therefore valuable for dissecting epigenetic contributions to proliferation, migration, and drug response, including sensitivity to H3K27 demethylase inhibitors.
Researchers can employ this polyclonal knockout for high-throughput screening of demethylase inhibitors, ChIP-qPCR profiling of H3K27me3 at gene promoters, RNA-seq analysis, and RT-qPCR quantification of CDKN1A and CDKN2A. Functional studies??proliferation, colony formation, and invasion assays??are readily performed, as are drug sensitivity tests. The cells also facilitate investigation of NOTCH-, TGF-??-, and NF-??B-driven inflammation and senescence. For further information, please contact Ascent Research.
