This product consists of a polyclonal population of ACOD1-knockout T-47D cells, generated by CRISPR/Cas9-mediated gene disruption to ablate aconitate decarboxylase 1 expression. The heterogeneous knockout pool provides a loss-of-function model for studying ACOD1-dependent processes without the clonal selection biases inherent in monoclonal lines, making it suitable for population-level analyses of immunometabolic functions in a breast cancer background. The targeted disruption is designed to eliminate the enzyme responsible for itaconate synthesis from cis-aconitate in the tricarboxylic acid cycle, thereby blocking production of this key immunometabolite and its downstream signaling effects.
The host cell line, T-47D, is a human breast carcinoma line originally isolated from the pleural effusion of a patient with infiltrating ductal carcinoma. This adherent epithelial line is positive for estrogen and progesterone receptors, modeling the luminal A molecular subtype of breast cancer and maintaining responsiveness to hormonal stimuli. Widely used in endocrinology and oncology, T-47D cells retain functional estrogen receptor signaling and are a standard platform for investigating hormone-dependent growth, therapeutic resistance, and epithelial-tumor microenvironment interactions.
ACOD1 encodes aconitate decarboxylase 1, the biosynthetic enzyme that converts cis-aconitate to itaconate, a metabolite with profound anti-inflammatory, antimicrobial, and immunomodulatory roles. Expression of ACOD1 is strongly induced by pro-inflammatory stimuli, including lipopolysaccharide, tumor necrosis factor, and interferons (IFN-??, IFN-??) acting through Toll-like receptor pathways. Once produced, itaconate alkylates cysteine residues on Kelch-like ECH-associated protein 1 (Keap1), leading to stabilization and nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) and subsequent transcription of antioxidant and cytoprotective genes. In parallel, itaconate inhibits succinate dehydrogenase (SDH), dampening mitochondrial reactive oxygen species and modulating macrophage polarization. Additional downstream effects include regulation of the NLRP3 inflammasome and activating transcription factor 3 (ATF3).
In the T-47D breast cancer context, disruption of ACOD1 and loss of itaconate production offer a powerful tool to dissect the intersection of immunometabolism and hormone receptor signaling. Emerging evidence links itaconate to tumor-associated inflammation and immune cell function within the tumor microenvironment. Because T-47D cells are estrogen receptor-positive, this knockout model enables investigation of potential crosstalk between itaconate-mediated Nrf2 activation and estrogen receptor pathways, and how metabolic reprogramming may influence hormone therapy responsiveness and immune surveillance. The polyclonal format supports studies where population heterogeneity more closely mimics physiological conditions.
Researchers can leverage the ACOD1-knockout T-47D polyclonal cells in a broad array of experimental applications. Representative assays include liquid chromatography?Cmass spectrometry for intracellular itaconate quantification, western blotting for Nrf2, Keap1, and ATF3, reverse transcription?Cquantitative PCR for ACOD1 transcript levels, metabolic flux analysis to trace TCA cycle perturbations, and cytokine profiling to assess inflammatory outputs. The model is particularly suited for cell proliferation, migration, and invasion assays in the context of breast cancer aggression, as well as estrogen receptor activity assays to test hormonal crosstalk. It is also valuable for tumor microenvironment reconstitution experiments and drug sensitivity screening. For additional details, please contact Ascent Research.
