The ACOD1 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of 786-O human renal epithelial cancer cells with targeted disruption of the ACOD1 gene, generating a loss-of-function model for immunometabolism research. This polyclonal product enables investigation of itaconate biosynthesis and its immunoregulatory roles, providing a robust tool for diverse experimental analyses. The genetic ablation of ACOD1 eliminates the enzymatic decarboxylation of cis-aconitate to itaconate, a metabolite with anti-inflammatory and antimicrobial properties.
The host cell line 786-O is a well-characterized model of clear cell renal cell carcinoma (ccRCC) harboring a loss-of-function mutation in the VHL tumor suppressor gene. This genetic background leads to constitutive stabilization of hypoxia-inducible factor 1-alpha (HIF-1??) even under normoxic conditions, resulting in aberrant transcriptional activation of genes involved in angiogenesis, glycolysis, and inflammation. 786-O cells exhibit typical ccRCC features, including high glycolytic flux and altered cytokine profiles, making them suitable for investigating metabolic and inflammatory interplay in kidney cancer. The VHL-HIF axis also intersects with immune signaling pathways, providing a relevant context for studying ACOD1 function.
ACOD1 (aconitate decarboxylase 1) catalyzes the conversion of the TCA cycle intermediate cis-aconitate to itaconate, a metabolite that exerts broad immunoregulatory effects. Itaconate inhibits succinate dehydrogenase (SDH) to reduce succinate-driven ROS and pro-inflammatory signaling, activates the Nrf2 transcription factor via alkylation of KEAP1, and suppresses the NLRP3 inflammasome, thereby dampening NF-??B-mediated cytokine production. Transcription of ACOD1 is induced by inflammatory stimuli such as LPS, TNF-??, IFN-??, and type I interferons through pathways involving NF-??B, IRF1, STAT1, and HIF-1??. Itaconate also modulates ATF3 and I??B??, further integrating metabolic and immune responses.
In VHL-deficient 786-O cells, constitutive HIF-1?? signaling likely drives ACOD1 expression, linking hypoxia responses to itaconate production. ACOD1 disruption allows investigation of the HIF-1??-driven immunometabolic network in tumor-associated inflammation and immune evasion. As ccRCC features aberrant cytokine secretion and immune infiltration, loss of itaconate may shift the balance toward pro-inflammatory pathways. This model thus facilitates examination of how cancer cells control innate immune signaling through endogenous metabolites.
These ACOD1 knockout polyclonal cells are suited for a range of functional assays, including Western blotting and RT-qPCR to confirm gene disruption and assess downstream targets, LC-MS-based itaconate quantification, multiplex cytokine profiling, NF-??B reporter assays, and Seahorse metabolic flux analysis to evaluate SDH inhibition and mitochondrial respiration. The model supports immunometabolism research, cancer inflammation studies, macrophage polarization analyses, and screening of itaconate-based therapeutic compounds. For further technical details, pricing, and availability, please contact Ascent Research.
