The KMT2C Knockout HT29 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma cell line, engineered to disrupt the KMT2C gene. This cell population provides a loss-of-function model for investigating KMT2C-dependent biological processes, including enhancer-mediated transcriptional regulation and tumor suppression. The polyclonal format retains genetic heterogeneity inherent to the parental line while introducing targeted gene disruption, enabling robust functional studies without clonal selection artifacts. Researchers can employ these cells in a wide range of assays to dissect KMT2C’s role in chromatin remodeling and colorectal cancer biology.
The host HT29 cell line is a well-established model of colorectal adenocarcinoma with an epithelial morphology. These cells harbor oncogenic BRAF V600E, APC, and TP53 mutations, alongside microsatellite instability, rendering them a clinically pertinent platform for colorectal cancer research. HT29 cells are extensively used to examine intestinal epithelial biology, including mucus secretion, barrier integrity, and tumorigenic signaling pathways. Their mutational landscape and MSI status make them particularly relevant for evaluating genotype-specific therapeutic vulnerabilities and molecular mechanisms underlying colorectal cancer progression.
KMT2C encodes a histone-lysine N-methyltransferase that catalyzes monomethylation of histone H3 at lysine 4 (H3K4me1), predominantly at enhancer regions, acting as a transcriptional coactivator. It is a core component of the COMPASS-like complex, interacting with ASH2L, RBBP5, WDR5, and DPY30, as well as PTIP, PA1, and NCOA6. In the Wnt/??-catenin pathway, KMT2C is recruited by the ??-catenin/TCF transcription factor complex to prime enhancers for activation, thereby regulating target genes involved in development and differentiation. Upstream regulators include FOXA1 and estrogen receptor, while downstream targets encompass enhancer-regulated genes, cell cycle regulators, and HOX loci.
The disruption of KMT2C in the HT29 background is particularly significant given the frequent mutation of this gene in colorectal tumors and its proposed tumor suppressor functions. HT29 cells carry constitutive activation of Wnt signaling due to APC mutation, creating a context in which KMT2C-mediated enhancer priming may be critical for modulating oncogenic gene expression. This knockout model enables dissection of how loss of KMT2C cooperates with existing driver mutations to alter chromatin landscapes, gene expression programs, and cellular phenotypes, offering insights into the mechanisms of enhancer dysfunction in colorectal cancer.
This polyclonal knockout cell population supports diverse research applications, including investigation of KMT2C’s tumor suppressor activity, enhancer biology, and synthetic lethal interactions. Key experimental readouts include western blotting and RT-qPCR to confirm target disruption, ChIP-qPCR for H3K4me1 occupancy at enhancers, RNA-seq for transcriptomic profiling, and functional assays such as cell proliferation, colony formation, and drug dose-response assessment. These cells facilitate screening for genotype-specific drug sensitivities and can be integrated into colorectal cancer progression models. For additional technical details or ordering information, please contact Ascent Research.
