The IMPA2 Knockout SK-HEP-1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal cell population with targeted disruption of the IMPA2 gene. This heterogeneous knockout pool, generated via CRISPR/Cas9-mediated gene disruption, serves as a loss-of-function model for inositol monophosphatase 2. The polyclonal format retains genetic diversity within the edited population, providing a robust system that avoids clonal bias for representative studies of IMPA2-dependent phenotypes.
The host SK-HEP-1 line is a human liver adenocarcinoma cell line derived from ascites fluid of a patient with liver adenocarcinoma. These adherent epithelial cells are a well-established model for hepatocellular carcinoma and display endothelial-like characteristics, making them valuable for tumor biology, angiogenesis, and metastasis research. Their reliable culture and compatibility with high-content imaging support diverse functional assays.
IMPA2 encodes an inositol monophosphatase that hydrolyzes inositol monophosphate to free myo-inositol and phosphate, sustaining myo-inositol pools essential for phosphatidylinositol (PtdIns) synthesis. In the phosphatidylinositol signaling system, IMPA2 acts upstream of PtdIns-4,5-bisphosphate (PIP2) regeneration, which is phosphorylated by PIP5K. Phospholipase C (PLC) then cleaves PIP2 into inositol-1,4,5-trisphosphate (IP3) and diacylglycerol (DAG). IP3 triggers calcium release via IP3 receptors, while DAG activates protein kinase C (PKC). IMPA2 is inhibited by lithium and valproic acid and transcriptionally regulated by SP1 and AP-2. Its knockout depletes free myo-inositol, impairing PIP2 resynthesis and attenuating IP3-mediated calcium mobilization and DAG-driven PKC signaling. IMPA2 interacts with IMPA1 and calbindin, linking inositol recycling to calcium homeostasis.
In SK-HEP-1 hepatocellular carcinoma cells, IMPA2 knockout provides a targeted tool to dissect inositol metabolism’s role in liver cancer pathology. Aberrant phosphatidylinositol signaling is implicated in hepatic tumor proliferation, survival, and migration, allowing this model to investigate metabolic dependencies and lithium response variability. The endothelial-like phenotype further enables studies of IMPA2 in tumor angiogenesis signaling.
This polyclonal knockout model supports applications including lithium mechanism studies, screening of inositol phosphate pathway modulators, and calcium signaling elucidation in cancer. Typical assays comprise inositol-1-phosphatase activity measurements, myo-inositol quantification by mass spectrometry, calcium flux imaging, and phospho-PKC substrate western blotting. Functional analyses like PIP2/IP3 turnover ELISA, lithium-treated cell viability, and migration/invasion assays provide translational insights. For technical inquiries, contact Ascent Research.
