The KDM5B Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population generated by disruption of the KDM5B gene in the HT29 human colorectal adenocarcinoma cell line. This product provides a loss-of-function model for studying KDM5B-mediated epigenetic regulation and its roles in colorectal cancer biology. The polyclonal format ensures a heterogeneous population of knockout cells suitable for functional studies without clonal selection bias.
HT29 is an established human colorectal adenocarcinoma epithelial cell line widely used as a model of intestinal epithelium. These cells exhibit an epithelial morphology, express differentiation markers such as villin, and form functional tight junctions. Importantly, HT29 cells harbor activating BRAF V600E and mutant TP53 mutations, which mimic common oncogenic drivers in colorectal cancer. This genetic background makes the line particularly relevant for investigating tumor suppressor pathways, drug responses, and epithelial biology in a colorectal context.
KDM5B encodes a histone H3 lysine 4 demethylase that specifically removes methyl groups from di- and trimethylated H3K4, leading to transcriptional repression of target genes. In the signaling network, KDM5B is regulated by oncogenic transcription factors such as MYC and E2F, as well as by HIF1A and miR-137. It interacts with co-repressor complexes including SIN3A, HDAC1, NuRD, and REST. By demethylating H3K4me3 at promoters of tumor suppressors like CDKN1A (p21) and CDKN2A (p16INK4a), KDM5B represses their expression, thereby promoting cell cycle progression and inhibiting senescence. This demethylase activity positions KDM5B as a key node in epigenetic control of proliferation and differentiation.
In the HT29 colorectal cancer model, KDM5B knockout disrupts this repressive epigenetic mechanism, potentially leading to derepression of CDKN1A and CDKN2A and subsequent cell cycle arrest or senescence. Given the BRAF V600E and TP53 mutant context, loss of KDM5B may alter sensitivity to targeted therapies or chemotherapeutics. This knockout system thus enables dissection of KDM5B-dependent transcriptional programs and interactions with other chromatin modifiers, providing insight into how epigenetic dysregulation contributes to colorectal tumorigenesis and maintenance.
Researchers can employ these polyclonal KDM5B knockout HT29 cells in a range of assays, including Western blotting for KDM5B and H3K4me3 levels, RT-qPCR for downstream targets CDKN1A and CDKN2A, and ChIP-qPCR to examine H3K4me3 enrichment changes. Functional studies such as cell proliferation, colony formation, flow cytometry-based cell cycle analysis, and apoptosis assays can reveal the impact of KDM5B loss. Moreover, drug sensitivity screening and RNA-seq experiments can uncover broader epigenetic and transcriptomic consequences. For additional information or technical inquiries, please contact Ascent Research.
