The KCNJ2 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting the human KCNJ2 gene in SK-HEP-1 liver adenocarcinoma cells. These cells contain a diverse array of indel mutations at the target locus, generated by non-homologous end joining, leading to functional gene disruption without single-cell cloning. This polyclonal pool allows robust population-level analyses of KCNJ2 deficiency.
SK-HEP-1 is a human liver adenocarcinoma cell line with both epithelial and endothelial features, expressing markers such as von Willebrand factor and CD34. Originally isolated from a hepatic adenocarcinoma, SK-HEP-1 has been employed in studies ranging from xenobiotic metabolism to endothelial cell biology, offering a robust platform for functional genomics. Its tumorigenic nature also facilitates cancer-relevant phenotypic screens, making it an ideal host for studying endothelial-like behaviors in a cancer context.
KCNJ2 encodes the inward rectifier potassium channel Kir2.1, which stabilizes resting membrane potential and controls excitability. Kir2.1 activity is modulated by PIP2, PKA, PKC, pH, Mg2+, and polyamines, and it forms complexes with DLG1, SNTA1, CAV3, and KCNJ4. Downstream, Kir2.1 regulates membrane voltage, L-type calcium channels, and Na+/K+ ATPase, thereby influencing calcium signaling and cellular effectors. Knockout is expected to depolarize the cell, alter calcium dynamics, and disrupt proliferation, migration, and angiogenesis pathways.
Given SK-HEP-1’s dual phenotype, KCNJ2 knockout provides a unique model to interrogate the intersection of ion channel activity and endothelial-like functions in cancer. The loss of Kir2.1-mediated currents is anticipated to perturb intracellular calcium homeostasis and downstream signaling cascades involving kinases and transcription factors that drive cell motility and angiogenesis, thereby offering a system to dissect these pathways in a hepatic tumor context.
Applications include patch-clamp electrophysiology, membrane potential assays, calcium imaging, and western blotting to confirm Kir2.1 loss. The model supports proliferation, migration, and tube formation assays to assess angiogenic potential. Further functional studies can include RT-qPCR and immunofluorescence to monitor KCNJ2 knockdown efficiency and compensatory changes in related channels. The cells are compatible with high-throughput screening platforms for ion channel modulators and co-culture experiments to probe tumor microenvironment interactions. For further information or to place an order, please contact Ascent Research.
