HCAR2 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma cell line, featuring targeted disruption of the HCAR2 gene. This polyclonal pool comprises a heterogeneous mixture of cells with loss-of-function mutations, reducing clonal bias and enabling robust population-level studies. The knockout model facilitates functional analysis of HCAR2-mediated signaling in colorectal cancer research.
The parental HCT 116 line is a male, near-diploid colorectal carcinoma epithelial model with MSI-high status and a KRAS G13D mutation, while retaining wild-type TP53. Its tumorigenic properties and well-defined genetic background make it a widely used system for investigating oncogenic pathways and therapeutic responses. The adherent growth and epithelial morphology support various phenotypic assays, including cell migration, proliferation, and apoptosis.
HCAR2 encodes a Gi-coupled receptor activated by niacin, butyrate, and ??-hydroxybutyrate. Upon activation, it inhibits adenylyl cyclase, reducing intracellular cAMP and PKA activity. This promotes PI3K/Akt signaling, which can attenuate NF-??B through phosphorylation of I??B??, while also modulating MAPK/ERK cascades. HCAR2 additionally regulates tight junction proteins such as occludin and claudin-1, essential for intestinal barrier function. Key interacting partners include G??i, G?¦?, ??-arrestin1/2, and GRK2/3, which mediate receptor desensitization and diverse downstream effects.
In HCT 116 cells, HCAR2 disruption may impair the anti-inflammatory and tumor-suppressive actions of butyrate and niacin. Butyrate, a gut microbiota-derived short-chain fatty acid, signals through HCAR2 to strengthen tight junctions and suppress NF-??B-driven inflammation. Ablation of HCAR2 in this MSI-high, KRAS-mutant background helps dissect the crosstalk between oncogenic signaling and nutrient-sensing pathways. This model is relevant for colorectal cancer, inflammatory bowel disease, and the tumor microenvironment, where HCAR2 influences epithelial proliferation and immune interactions.
Applications span cancer biology, anti-inflammatory signaling, intestinal barrier research, metabolic regulation, and drug target validation. The polyclonal knockout pool is compatible with cAMP assays, NF-??B reporter assays, western blotting for p-Akt and ERK1/2, and Transwell migration assays. RT-qPCR and immunofluorescence enable confirmation of target disruption and downstream gene analysis. Researchers can employ this model to investigate HCAR2??s roles in atherosclerosis, dyslipidemia, or metabolic syndrome, and to screen HCAR2-targeting compounds. For further information or technical assistance, please contact Ascent Research.
