The KCNK1 Knockout NCI-H1975 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population derived from NCI-H1975 human lung adenocarcinoma cells, with targeted disruption of the KCNK1 tumor suppressor gene. This polyclonal pool retains the genetic diversity of the parental line, minimizing artifacts associated with single-cell cloning and enabling robust functional studies such as pooled viability screens, drug sensitivity profiling, and signaling analysis.
The NCI-H1975 host cell line, isolated from the pleural effusion of a never-smoker female patient, harbors EGFR L858R and T790M mutations??the latter conferring resistance to first-generation EGFR tyrosine kinase inhibitors (TKIs). Consequently, it is a widely employed model for investigating EGFR-driven oncogenic signaling and acquired TKI resistance in non-small cell lung cancer.
KCNK1 encodes TWIK-1, a two-pore domain potassium channel that mediates background potassium currents to maintain resting membrane potential. Its activity is regulated by EGFR/MAPK signaling, hypoxia, miR-7-5p, and post-translational modifications including PKA/PKC phosphorylation and PIAS3-catalyzed SUMOylation. TWIK-1 interacts with 14-3-3 adaptors, heterodimerizes with K2P channels TASK-1 and TREK-1, and associates with ??-arrestins and the actin cytoskeleton. When KCNK1 is disrupted, membrane depolarization triggers calcium influx, which activates the AKT/mTOR pathway. This leads to increased Bcl-2, decreased Bax, and reduced caspase-3 activity, inhibiting apoptosis, while simultaneously upregulating cyclin D1 and downregulating p21 to drive cell cycle progression. Therefore, KCNK1 operates as a tumor suppressor, and its loss promotes proliferation and survival.
Within this EGFR-mutant background, KCNK1 loss is predicted to further stimulate the AKT/mTOR axis, leading to enhanced cell proliferation, apoptosis suppression, and possible changes in TKI susceptibility. This polyclonal knockout model thus provides a powerful system to elucidate how TWIK-1 channel dysfunction intersects with EGFR-mediated oncogenic networks and to validate the functional significance of KCNK1 as a tumor suppressor in lung adenocarcinoma.
This product supports diverse experimental approaches. Western blotting and RT-qPCR can confirm KCNK1 ablation and monitor downstream AKT, phospho-AKT, Bcl-2, and Bax levels. Electrophysiological recordings measure TWIK-1 currents and membrane potential changes. Cell-based assays include viability (MTT/CellTiter-Glo), apoptosis (Annexin V/PI), cell cycle flow cytometry, and Transwell migration/invasion tests. Transcriptomic analysis by RNA-seq reveals global expression changes, and in vivo tumorigenicity studies assess the impact of KCNK1 loss on tumor growth. For further inquiries, contact Ascent Research.
