The ACOD1 Knockout SK-HEP-1 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human liver adenocarcinoma-derived endothelial cell line. This polyclonal pool consists of cells harboring disruptions in the ACOD1 gene, enabling loss-of-function studies of aconitate decarboxylase (IRG1) without artifacts of clonal selection. The polyclonal format better represents the genetic heterogeneity of a native cell population while achieving effective target-gene disruption.
SK-HEP-1 cells serve as a widely accepted model of liver sinusoidal endothelial cells (LSECs), originally established from a human hepatic adenocarcinoma. These cells retain key phenotypic features of LSECs, including the capacity for filtration, endocytosis, and immune regulatory functions. Their endothelial character and hepatic origin make them particularly valuable for investigating vascular biology, drug metabolism, and the liver??s immune microenvironment.
ACOD1 encodes aconitate decarboxylase, the enzyme responsible for catalyzing the production of itaconate from the TCA cycle intermediate cis-aconitate. In response to inflammatory signals such as LPS acting through TLR4, or cytokines including IFN-?? and TNF-??, ACOD1 transcription is robustly upregulated via the coordinated activity of transcription factors NF-??B, STAT1, and IRF1. The resultant itaconate exerts broad anti-inflammatory effects: it competitively inhibits succinate dehydrogenase (SDH), leading to succinate accumulation; it alkylates KEAP1, activating the transcription factor Nrf2 to drive antioxidant gene expression; and it directly blocks NLRP3 inflammasome activation, thereby suppressing IL-1?? maturation. Consequently, ACOD1 acts as a critical rheostat in innate immune signaling cascades.
In the context of liver sinusoidal endothelial cells, ACOD1-mediated itaconate production may play a pivotal role in maintaining hepatic immune homeostasis. LSECs are strategically positioned to sense blood-borne pathogens and inflammatory mediators, and they can modulate the recruitment and activation of immune cells. Disruption of ACOD1 in SK-HEP-1 cells allows researchers to dissect how loss of itaconate synthesis alters endothelial responses to TLR4 agonists, cytokines, and bacterial challenge. This knockout model thus provides a unique tool to explore the cell-autonomous contributions of ACOD1 to LSEC-driven inflammation, metabolic stress, and immune tolerance within the hepatic sinusoid.
These polyclonal knockout cells are well suited for advanced experimental techniques including LC-MS quantification of itaconate, Seahorse metabolic flux analysis, cytokine profiling via ELISA or Luminex, RT-qPCR for ACOD1, IL-1??, and IL-6, Western blotting for Nrf2, KEAP1, and NLRP3, SDH activity assays, NF-??B luciferase reporter assays, and bacterial co-culture systems. For further details and ordering information, please contact Ascent Research.
