This product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HCT 116 colorectal carcinoma cell line, engineered to disrupt the KDM5C gene. The polyclonal pool harbors a heterogeneous mixture of loss-of-function alleles generated by non-homologous end joining, providing a robust tool for studying KDM5C-dependent epigenetic regulation. As a mixed population, it captures diverse mutational outcomes and is ideal for pooled functional genomics, drug screening, and bulk biochemical assays, offering a cost-effective alternative to single-cell-derived clones while preserving biological complexity. The knockout is validated at the genomic level to confirm target-gene disruption, enabling reliable investigation of KDM5C??s role in chromatin biology and cancer.
HCT 116 is a widely employed model of human colorectal adenocarcinoma featuring a KRAS G13D mutation, a PIK3CA H1047R activating mutation, and MLH1 deficiency leading to microsatellite instability. These oncogenic alterations drive constitutive MAPK and PI3K/AKT signaling, creating a permissive context for tumor cell proliferation and survival. The epithelial origin and rapid growth kinetics of HCT 116 cells make them particularly suitable for high-throughput drug sensitivity assays, xenograft tumorigenesis studies, and exploring cancer epigenetics. Their well-characterized genetic background and established use in colorectal cancer research provide a reproducible platform for dissecting the functional consequences of KDM5C loss.
KDM5C encodes a histone H3K4me3/me2 demethylase that acts as a transcriptional repressor by removing activating methyl marks from promoter regions. As part of multiprotein corepressor complexes, KDM5C interacts with HDAC1/2 and SIN3A to coordinate histone deacetylation and demethylation, and it is recruited by transcription factors such as REST/NRSF and E2F1 to silence specific gene programs. KDM5C is negatively regulated by miR-137 and can be targeted for proteasomal degradation, while its own activity directly represses key cell cycle and oncogenic targets including CCND1, CCNE1, CDKN1A, MYC, and HOX genes. Through these interactions, KDM5C modulates the balance between H3K4me3-marked active chromatin and silenced states, influencing cell proliferation, differentiation, and tumor suppression.
Disruption of KDM5C in HCT 116 cells abolishes its demethylase function, resulting in hypermethylation of H3K4me3 at promoter regions and aberrant transcriptional activation of oncogenic and cell cycle-promoting genes. This epigenetic rewiring enhances proliferative drive, accelerates cell cycle progression, and amplifies tumorigenic potential, consistent with KDM5C??s tumor-suppressive roles in certain contexts. The combined effect of KRAS and PIK3CA mutations together with loss of KDM5C-mediated repression creates a synthetic vulnerability that can be exploited to screen for histone demethylase inhibitors or other chromatin-modifying therapies. Thus, this knockout model provides a physiologically relevant system for studying colorectal cancer epigenetics and testing therapeutic interventions.
This KDM5C polyclonal knockout cell pool is ideally suited for a wide range of research applications, including chromatin immunoprecipitation followed by quantitative PCR (ChIP-qPCR) to assess H3K4me3 enrichment at specific promoters, RNA-sequencing for transcriptome-wide profiling of KDM5C-dependent gene regulation, and western blotting to confirm target protein levels. Cell-based assays such as MTT proliferation measurements and colony formation tests reliably quantitate growth phenotypes, while flow cytometry enables cell cycle analysis. In vivo xenograft models can evaluate tumor growth modulation, and drug sensitivity screening can identify compounds that selectively target KDM5C-deficient cells. For further technical details, please contact Ascent Research.
