The IREB2 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell pool designed for functional studies of the IREB2 gene. This heterogeneous population of SK-HEP-1 cells harbors targeted disruption of IREB2, enabling investigation of iron regulatory protein 2 (IRP2) in a mixed knockout background without clonal selection. The pool is optimized for a range of molecular and cellular assays.
The host SK-HEP-1 cell line originates from human hepatic endothelial cells isolated from the ascitic fluid of an adenocarcinoma patient. Retaining endothelial traits, SK-HEP-1 serves as a model for liver sinusoidal endothelial cells, which mediate blood filtration, nutrient transport, endocytosis, and metabolic regulation, and contribute to hepatic immune tolerance. This background provides a physiologically relevant platform for studying liver iron homeostasis and its dysregulation.
IREB2 encodes IRP2, a key post-transcriptional regulator of cellular iron homeostasis. When intracellular iron is low, IRP2 binds iron-responsive elements (IREs) in target mRNAs, increasing stability of transferrin receptor (TFRC) mRNA to promote iron uptake and blocking translation of ferritin heavy chain (FTH1), ferritin light chain (FTL), ferroportin (SLC40A1), and other transcripts to limit iron storage/export. IRP2 activity is suppressed under iron-replete or high-oxygen conditions via FBXL5-mediated ubiquitination and proteasomal degradation. Additional upstream cues include nitric oxide and hypoxia-inducible factor 2?? (HIF-2??), while downstream effects extend to ALAS2 and SLC11A2. IRP2 interacts with iron-sulfur cluster biosynthetic factors ISCU and NFS1, and functionally overlaps with IRP1.
In SK-HEP-1 cells, IREB2 knockout disrupts iron regulation within hepatic endothelial cells, offering a model to explore iron-handling defects relevant to hemochromatosis, iron-loading anemias, and liver fibrosis. The link between IRP2 and HIF-2?? further allows dissection of cross-talk between iron status and hypoxic signaling in the liver microenvironment. This model is especially useful for evaluating how iron dysregulation influences endothelial cell function in the liver sinusoid.
Typical applications include western blotting, RT-qPCR, RNA immunoprecipitation, cellular iron uptake assays, flow cytometry for TFRC surface levels, and IRE-luciferase reporter measurements. The polyclonal pool is suitable for drug screens targeting iron chelators or FBXL5 activators and for studies on cancer iron addiction and neurodegenerative disorders linked to brain iron accumulation. For more details, please contact Ascent Research.
