The ACOD1 Knockout HCT 116 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human ACOD1 gene in the HCT 116 colorectal carcinoma cell line. This polyclonal format offers a pool of cells with heterogeneous gene-edited alleles, enabling functional loss-of-function studies without the need for clonal isolation. The product is intended for advanced biomedical research applications in immunometabolism, inflammation, and cancer biology.
HCT 116 is a well-characterized human colon carcinoma cell line exhibiting epithelial morphology and high microsatellite instability (MSI-H), harboring a KRAS G13D driver mutation while retaining wild-type TP53. This cell line serves as a widely used model for colon cancer research, particularly for exploring mechanisms of tumorigenesis, therapeutic responses, and immune evasion. The MSI-H status renders HCT 116 susceptible to frameshift mutations and contributes to its distinct mutational landscape, making it a relevant system for studying genomic instability.
ACOD1 (aconitate decarboxylase 1) catalyzes the conversion of cis-aconitate to itaconate, a metabolite linking the TCA cycle to immune regulation. Upon stimulation by upstream cues such as LPS, TNF??, and type I interferons, ACOD1 is upregulated, leading to itaconate accumulation. Itaconate exerts anti-inflammatory effects by inhibiting succinate dehydrogenase (SDH), alkylating KEAP1 to activate the NRF2 antioxidant pathway, and suppressing NLRP3 inflammasome assembly. ACOD1 also interacts with ATF3, I??B??, and GAPDH, positioning it at a critical node connecting metabolic reprogramming, oxidative stress responses, and innate immune signaling.
In the HCT 116 colorectal cancer background, ACOD1 disruption is particularly relevant for dissecting the crosstalk between metabolic alterations and inflammation-driven tumor progression. The KRAS-mutant, MSI-H genotype of HCT 116 creates a pro-inflammatory tumor microenvironment in which ACOD1-derived itaconate may modulate NF-??B activity, cytokine production, and anti-tumor immune responses. By eliminating itaconate synthesis, this knockout model allows researchers to investigate the impact of abolished SDH inhibition and reduced NRF2 activation on colorectal cancer cell survival, proliferation, and susceptibility to immune cell-mediated killing.
Typical applications include investigating the role of itaconate in colorectal cancer inflammation and metabolic reprogramming. Researchers can employ this model in Western blot analyses of ACOD1 and downstream targets, LC-MS measurement of itaconate levels, SDH enzymatic activity assays, and NRF2 immunofluorescence. Functional assays such as NF-??B luciferase reporters, multiplex cytokine profiling via ELISA, cell viability, and apoptosis measurements are readily compatible. This polyclonal knockout cell pool provides a flexible and powerful tool for pathway interrogation and drug screening. For further technical details and ordering information, please contact Ascent Research.
