The ACOD1 Knockout AGS Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout population derived from human AGS gastric adenocarcinoma cells. ACOD1 (immunoresponsive gene 1, IRG1) encodes aconitate decarboxylase 1, the rate-limiting enzyme for itaconate biosynthesis from the TCA cycle intermediate cis-aconitate. This polyclonal format, generated by non-clonal pool selection, provides a heterogeneous genetic background that reflects natural population diversity and allows robust phenotypic comparisons with wild-type AGS cells.
AGS cells, isolated from a gastric adenocarcinoma, are widely employed as an in vitro model of gastric epithelium. They exhibit key features of epithelial cells, including mucin secretion and tight junction formation, and retain functional signaling networks implicated in gastric inflammation, Helicobacter pylori infection, and carcinogenesis. Consequently, they offer a physiologically relevant platform to investigate the roles of metabolic enzymes in gastric mucosal immunity and cancer progression.
ACOD1 directs metabolic flux toward itaconate, a metabolite with potent immunomodulatory properties. Itaconate inhibits succinate dehydrogenase (SDH) activity, triggering succinate accumulation and pseudohypoxia-like responses, while concurrently activating the Nrf2?CKEAP1 antioxidant axis and suppressing NF-??B-driven transcription and NLRP3 inflammasome assembly. ACOD1 transcription is itself regulated by inflammatory stimuli: NF-??B, IRF1, and STAT1 bind its promoter upon exposure to LPS, TNF, or IFN-??. This multi-layered regulation positions ACOD1 as a critical nexus between cellular metabolism and innate immune responses.
Disruption of ACOD1 in AGS gastric cancer cells eliminates itaconate production, thereby altering metabolic and inflammatory pathways. The loss of itaconate is predicted to relieve SDH inhibition, diminish Nrf2-mediated cytoprotective gene expression, and enhance NLRP3 inflammasome and NF-??B activity, leading to increased secretion of pro-inflammatory cytokines. This model is particularly relevant for studying the interplay between metabolic reprogramming and inflammation in gastric carcinogenesis and for evaluating epithelial responses to bacterial pathogens such as H. pylori.
Typical experimental applications include western blotting and RT-qPCR for protein and transcript analysis, itaconate level determination by mass spectrometry or enzymatic assays, NF-??B reporter assays, NLRP3 inflammasome activation studies, multiplex cytokine profiling, SDH enzymatic activity measurements, Nrf2 target gene expression analysis, and functional assays for cell migration, invasion, and drug sensitivity. Researchers in gastric cancer, inflammatory bowel disease, sepsis, and immunometabolism will find this model valuable for mechanistic studies and drug target validation. For further information, please contact Ascent Research.
