The ACOD1 Knockout Ca Ski Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the ACOD1 (IRG1) gene in a human cervical carcinoma background. This reagent provides a heterogeneous pool of gene-disrupted cells, enabling functional interrogation of itaconate biosynthesis and its downstream immunomodulatory networks without clonal selection artifacts. As a polyclonal knockout model, it is suitable for experiments where population-level responses are prioritized, such as signaling pathway analysis and metabolic profiling.
The host Ca Ski cell line was originally established from a metastasis of a cervical squamous cell carcinoma and harbors an integrated human papillomavirus type 16 (HPV-16) genome. This adherent epithelial cell line is widely employed in cancer research, particularly for studying HPV-driven oncogenesis, tumor cell signaling, and cervical cancer biology. The presence of viral oncoproteins E6 and E7, which inactivate p53 and Rb, respectively, makes Ca Ski a valuable model for exploring virus?Chost interactions and the molecular underpinnings of cervical carcinoma progression.
ACOD1 encodes cis-aconitate decarboxylase, which converts cis-aconitate to itaconate, a metabolite with potent immunoregulatory and antibacterial properties. Itaconate inhibits succinate dehydrogenase (SDH), alkylates KEAP1 to activate Nrf2-dependent antioxidant responses, and blocks NLRP3 inflammasome activation. ACOD1 is transcriptionally regulated by NF-??B and IRF1 downstream of LPS, TNF??, and type I interferon receptors. Key downstream mediators include ATF3 and the Nrf2 pathway. Disrupting ACOD1 in Ca Ski cells thereby impairs itaconate production and perturbs these interconnected signaling axes.
In Ca Ski cells, which exhibit constitutive NF-??B activity driven by HPV oncoproteins, ACOD1 knockout provides a system to dissect crosstalk between viral pathogenesis and immunometabolism. Researchers can evaluate how loss of itaconate synthesis influences HPV-mediated inflammatory responses, oxidative stress defense, and tumor cell viability. As a standard cervical carcinoma model, these knockout cells are relevant for studying the succinate?Citaconate?CSDH axis in cancer metabolism and exploring ACOD1 as a target in HPV-associated malignancies.
Applications include investigation of itaconate-mediated Nrf2 activation using western blotting and luciferase reporter assays, NLRP3 inflammasome functional analyses by caspase-1 activation and IL-1?? release, metabolic flux studies with LC-MS-based itaconate quantification, and SDH enzymatic activity measurements following inflammatory stimulation. The polyclonal population offers robust and scalable material for high-throughput screening or deep mechanistic studies. For further information, please contact Ascent Research.
