The ACOD1 Knockout HL-60 Polyclonal Cells are a ready-to-use population of HL-60 cells that have undergone CRISPR/Cas9-mediated gene disruption at the ACOD1 locus. As a polyclonal knockout product, this population comprises a heterogeneous mixture of edited cells without single-cell cloning, providing a robust loss-of-function model for studying ACOD1-dependent processes. This format avoids clonal selection artifacts and is well-suited for experiments where biological variability is acceptable or for initial functional screens.
The HL-60 cell line is derived from human acute promyelocytic leukemia and is p53-null, capable of differentiating into granulocyte- or macrophage-like cells upon stimulation with DMSO or PMA, respectively. This cell line is widely employed as a model for myeloid differentiation and innate immune function, gaining phagocytic and respiratory burst activity upon maturation. Its genetic tractability and well-characterized intracellular signaling pathways make HL-60 an ideal host for generating knockouts to dissect gene functions in inflammatory responses and immunometabolism.
ACOD1 (IRG1) catalyzes the decarboxylation of cis-aconitate to itaconate, a metabolite that accumulates in activated macrophages. Its expression is strongly induced by LPS, IFN-??, TNF, and IL-1??, primarily via transcription factors NF-??B, STAT1, IRF1, and HIF-1??. Downstream, itaconate inhibits succinate dehydrogenase (SDH), leading to TCA cycle remodeling and succinate accumulation. Itaconate also alkylates cysteine residues on KEAP1, resulting in NRF2 stabilization and activation of antioxidant genes, and concurrently induces ATF3 and suppresses NLRP3 inflammasome assembly. Additionally, itaconate directly inhibits bacterial isocitrate lyase, exerting antimicrobial effects. Interacting partners include ACO2 and KEAP1. Thus, ACOD1 functions as a pivotal immunometabolic regulator, linking innate immune signaling with cellular metabolism and host defense.
In the HL-60 context, ACOD1 knockout permits dissection of endogenous itaconate’s contribution to NF-??B pathway modulation, NRF2 activation, and inflammasome control during myeloid differentiation. The ability to chemically induce macrophage-like differentiation allows researchers to study ACOD1’s role in antimicrobial defense mechanisms, such as itaconate-mediated inhibition of bacterial isocitrate lyase. Furthermore, the p53-null status of HL-60 cells provides a unique opportunity to explore p53-independent metabolic reprogramming pathways triggered by inflammatory stimuli. This model thus offers a simplified yet physiologically relevant platform to investigate itaconate-dependent immunomodulation.
The ACOD1 Knockout HL-60 Polyclonal Cells enable knockout validation by RT-qPCR and western blotting, LC?CMS quantification of itaconate, and ELISAs for TNF, IL-6, IL-1??. Signaling can be examined with NF-??B and NRF2 reporter assays. Differentiation with PMA/DMSO coupled with flow cytometry assesses macrophage markers and antimicrobial function. These cells are ideal for screening immunometabolic compounds and studying inflammation-driven metabolic shifts. For further details, please contact Ascent Research.
