The KDM3B Knouckout HT29 Polyclonal Cells product consists of a pool of HT29 colorectal adenocarcinoma cells carrying a CRISPR/Cas9-mediated disruption of the KDM3B locus. This polyclonal population provides a loss-of-function model suitable for investigating KDM3B functions in a genetically diverse, non-clonal context. The gene-edited cells are generated without isolating single clones, preserving the inherent heterogeneity of the parental line while permitting robust analysis of target gene depletion in bulk-cell assays.
The HT29 cell line originates from a primary colorectal adenocarcinoma of a 44-year-old Caucasian female and harbors mutations in key oncogenes and tumor suppressors, including APC, TP53, and KRAS. These epithelial cells can differentiate into enterocyte-like phenotypes under defined culture conditions, serving as a versatile model for intestinal epithelial biology, colorectal cancer progression, and differentiation studies. Their genetic background closely mirrors the mutational landscape of sporadic colorectal tumors.
KDM3B encodes a histone demethylase that removes mono- and dimethyl groups from lysine 9 of histone H3 (H3K9me1/2), relieving transcriptional repression at target gene promoters. This enzyme is regulated by hypoxia-inducible factor 1-alpha (HIF1A) and androgen receptor signaling, with additional inputs from PI3K/AKT and MYC pathways. KDM3B-mediated demethylation activates transcription of key downstream targets such as CCND1, CDKN1A, NOTCH1, HES1, and c-MYC, linking epigenetic control to cell cycle regulation, differentiation, and oncogenic programs.
In HT29 cells, KDM3B knockout leads to increased H3K9me1/2 levels, causing transcriptional silencing of genes critical for proliferation, hypoxia adaptation, and Notch signaling. This disruption perturbs the balance between undifferentiated and differentiated states, which is central to colorectal tumorigenesis and therapeutic sensitivity. The polyclonal nature of the edited population enables examination of how KDM3B loss cooperates with endogenous APC, TP53, and KRAS mutations to drive aggressive phenotypes in a heterogeneous tumor cell environment.
This knockout model supports diverse experimental workflows, including western blotting, RT-qPCR, and ChIP-qPCR to quantify H3K9me2 changes and target gene expression. Functional assays such as proliferation, colony formation, and migration/invasion tests evaluate KDM3B??s role in tumor cell behavior. Transcriptomics via RNA-seq and drug sensitivity screens further elucidate downstream pathways and resistance mechanisms. The polyclonal format is optimal for pooled CRISPR screens and studies requiring biological variability. For further information, please contact Ascent Research.
