The ACOD1 Knockout HT29 Polyclonal Cells consist of a genetically heterogeneous polyclonal population of HT29 cells in which the ACOD1 gene has been disrupted using CRISPR/Cas9 technology. This product provides a loss-of-function model for studying the immune-responsive gene encoding aconitate decarboxylase 1 (IRG1), without reliance on a single clonal isolate. The polyclonal format reduces the risk of clonal artifacts and better represents the distribution of knockout-induced phenotypes across the cell population, making it suitable for robust and physiologically relevant in vitro investigations.
The HT29 parental cell line is a well-established human colorectal adenocarcinoma epithelial line derived from a primary tumor of a 44-year-old Caucasian female. HT29 cells retain epithelial morphology and are widely employed as a tumorigenic model for colorectal cancer biology, intestinal epithelial barrier function, and drug response studies. They have been characterized extensively for their ability to form xenograft tumors and to differentiate into enterocyte-like cells under appropriate conditions, providing a relevant background for interrogating gene function in intestinal malignancy and inflammation.
ACOD1 (IRG1) catalyzes the decarboxylation of cis-aconitate to produce itaconate, a metabolite connecting mitochondrial metabolism to innate immunity. Its expression is induced downstream of TLR4 by LPS, type I interferons, and the transcription factors NF-??B and IRF1, with additional modulation by STAT1 and TNF-??. Itaconate alkylates KEAP1 at cysteine residues, leading to NRF2 stabilization and activation of antioxidant defenses. Concurrently, itaconate inhibits succinate dehydrogenase and the NLRP3 inflammasome, suppressing pro-inflammatory cytokine production. These multiple targets position ACOD1 as a pivotal regulator at the crossroads of glycolysis, the TCA cycle, and inflammatory signaling.
In the HT29 colorectal cancer model, the ACOD1/itaconate axis is critical because chronic inflammation drives tumorigenesis. HT29 cells express TLR4 and respond to inflammatory stimuli, making them suitable for studying how itaconate synthesis affects NF-??B signaling, NRF2-mediated antioxidant responses, and metabolic reprogramming. ACOD1 knockout in HT29 thus enables investigation of tumor-intrinsic roles of itaconate and evaluation of targeting this metabolic pathway in colorectal cancer.
This knockout model supports diverse experimental applications. Co-culture with macrophages enables study of epithelial-derived itaconate effects on immune cell polarization. NF-??B reporter assays, RT-qPCR, and ELISA can quantify inflammatory responses, while LC-MS and Seahorse analysis provide metabolic profiling. Immunofluorescence for NRF2 localization and Western blotting for pathway components further validate signaling outcomes. For further information, please contact Ascent Research.
