The ACOD1 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to abolish expression of the human ACOD1 gene in the HAP1 near-haploid cell line. ACOD1 encodes aconitate decarboxylase, the enzyme responsible for catalyzing the production of the immunometabolite itaconate from the TCA cycle intermediate cis-aconitate. This polyclonal knockout pool provides a mixed population of edited cells and serves as a powerful loss-of-function tool for interrogating the role of the itaconate pathway in inflammatory signaling, metabolic reprogramming, and cellular stress responses without the limitations of clonal selection.
The host HAP1 cell line is a human fibroblast-like, adherent line originally derived from the KBM-7 chronic myeloid leukemia cell line. Its near-haploid karyotype makes it particularly amenable to gene editing and functional genomics studies, as the presence of a single copy of most genes facilitates efficient knockout generation and phenotypic analysis. HAP1 cells retain key signaling pathways relevant to cancer and metabolic research, and their robust growth characteristics and suitability for high-throughput screening further enhance their utility as a model system.
ACOD1 is rapidly induced upon inflammatory stimulation and sits at a critical node linking metabolism to immunity. Upstream activators such as LPS (via TLR4), TNF-??, and type I interferons converge on transcription factors NF-??B, IRF1, and STAT1 to drive ACOD1 expression. The resulting itaconate production mediates potent anti-inflammatory effects through at least two mechanisms: competitive inhibition of succinate dehydrogenase (SDH) to modulate mitochondrial respiration and alkylation of KEAP1, which releases the transcription factor NRF2 to induce cytoprotective gene programs including HO-1 and NQO1. In parallel, itaconate suppresses pro-inflammatory cytokines such as IL-1?? and IL-6, dampening the inflammatory response. Disruption of ACOD1 in the knockout cell pool thus abrogates these regulatory circuits, allowing researchers to dissect the contribution of itaconate to the anti-inflammatory and metabolic phenotypes controlled by NRF2 and SDH.
In the HAP1 background, loss of ACOD1 offers a defined platform to study the intersection of cancer biology and immunometabolism. Although HAP1 cells are not immune cells, they can be stimulated with inflammatory agonists or engineered to express macrophage markers, making them a tractable system for examining early signaling events downstream of ACOD1 without the complexity of primary cells. The polyclonal nature of the product avoids clonal artifacts while still enabling robust population-level analyses, and the near-haploid genome simplifies the interpretation of functional assays, as compensatory mechanisms are reduced. This model is especially valuable for dissecting how itaconate modulates metabolic flux and redox homeostasis, and for screening modulators of the KEAP1?CNRF2 axis.
This knockout cell product is suited for a broad range of experimental applications, including macrophage immunometabolism studies, inflammation research, infection models, cancer metabolism investigations, and drug target validation. Representative assays compatible with the model include western blotting to confirm loss of ACOD1 protein, liquid chromatography?Cmass spectrometry (LC-MS) to quantify itaconate levels, RT-qPCR for downstream inflammatory cytokines (e.g., IL-1??, TNF-??), NF-??B luciferase reporter assays, NRF2 activation assays (such as ARE luciferase), succinate dehydrogenase activity measurements, and immunofluorescence or flow cytometry for phenotypic characterization. For additional technical details or to discuss custom cell engineering projects, please contact Ascent Research.
