The ACOD1 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal HeLa cell population with targeted disruption of the ACOD1 gene. This polyclonal knockout model provides a heterogeneous pool of edited cells, suitable for functional studies without clonal selection, enabling robust assessment of aconitate decarboxylase 1 function. The product serves as a versatile loss-of-function tool for investigating itaconate biology.
The parental HeLa cell line is an immortalized human epithelial cell line from cervical adenocarcinoma, extensively characterized and widely used in cancer biology, virology, signal transduction, and metabolism research. Its well-documented genome and robust growth make it a reliable host for gene editing, particularly for examining metabolic and inflammatory signaling in an epithelial cancer context.
ACOD1 encodes aconitate decarboxylase 1, which converts cis-aconitate to itaconate, a key immunometabolite that inhibits succinate dehydrogenase (SDH) in the TCA cycle and alkylates Keap1 cysteines, activating Nrf2 and inducing antioxidant genes such as HMOX1 and NQO1. Itaconate also suppresses the NLRP3 inflammasome and reduces IL-6 production. ACOD1 expression is strongly induced by LPS, type I interferons, and TNF-?? through NF-??B, IRF1, STAT1, and IRF3. Downstream, itaconate modulates SDH, Nrf2 targets, NLRP3, and ferroptosis-related proteins, placing ACOD1 at the nexus of inflammation, oxidative stress, and cell death.
In HeLa epithelial cancer cells, ACOD1 knockout allows dissection of itaconate-dependent regulation of inflammatory and metabolic pathways outside immune cells. HeLa cells express NF-??B and JAK/STAT signaling components, enabling study of how ACOD1 disruption alters responses to LPS, TNF-??, or interferons. This model can probe links between the TCA cycle, Nrf2 activity, and ferroptosis susceptibility, relevant to itaconate’s emerging role in tumor survival, thus facilitating investigation of cell-type-specific functions of the itaconate pathway.
Applications include characterizing itaconate production and SDH activity by LC-MS metabolomics, assessing Nrf2 target gene induction by RT-qPCR, quantifying cytokine secretion by ELISA, and evaluating NLRP3 inflammasome activation. The model also supports ferroptosis studies via lipid peroxidation and cell viability assays, and screening for pathway modulators. Western blotting can monitor STAT1 phosphorylation and Keap1 modification. This knockout polyclonal cell population is an essential tool for immunometabolism, cancer metabolism, and host-pathogen research. For ordering or technical support, contact Ascent Research.
