The KCNJ2 Knockout KYSE-150 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from KYSE-150 human esophageal squamous cell carcinoma cells, featuring targeted disruption of the KCNJ2 locus. This heterogeneous pool of edited cells avoids clonal artifacts and is ideal for population-level studies of gene function, drug responses, and electrophysiological measurements in a cancer-relevant background. Gene disruption is verified at the population level, with allele variability expected.
The parental KYSE-150 cell line, originally established from a primary esophageal squamous cell carcinoma, is a widely used model of ESCC. These adherent epithelial cells endogenously express KCNJ2, which encodes the inward rectifier potassium channel Kir2.1, contributing to resting membrane potential and ion homeostasis. The knockout model enables direct functional analysis of Kir2.1 in a disease-relevant context.
KCNJ2 encodes Kir2.1, a strong inward rectifier potassium channel that stabilizes the resting membrane potential by mediating potassium influx at hyperpolarized potentials. Kir2.1 activity is regulated by PKA and PKC phosphorylation, GPCR signaling, and PIP2 binding, with loss of PIP2 causing channel inactivation. The channel forms homotetramers and interacts with scaffolding proteins DLG1, CASK, and SNTB2 for membrane localization. Downstream, Kir2.1 influences voltage-gated calcium channels and the ERK pathway, thereby affecting cell proliferation and apoptosis. This positions KCNJ2 as a critical link between ion transport and cellular signaling.
In KYSE-150 cells, KCNJ2 knockout disrupts potassium homeostasis and depolarizes membrane potential, potentially altering cell cycle progression and apoptotic sensitivity. This model is valuable for exploring how loss of Kir2.1 affects ESCC phenotypes, given emerging evidence that ion channels modulate cancer cell behavior. Moreover, KCNJ2 mutations are associated with Andersen-Tawil syndrome and cardiac arrhythmias, underscoring its broad physiological relevance and making this model applicable to both cancer and cardiac research.
Typical applications include patch-clamp electrophysiology to characterize potassium current defects, fluorescent membrane potential assays for compound screening, and functional assessments via MTT/CCK-8 proliferation, Annexin V apoptosis, and invasion assays. Validation can be performed using western blotting for Kir2.1 and RT-qPCR for KCNJ2 mRNA, with transcriptome-wide analysis via RNA-seq. These tools facilitate investigation of ion channel roles in esophageal squamous cell carcinoma and drug testing for ion channel-related diseases. For further information, contact Ascent Research.
