The KCNK3 Knockout T-47D Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population generated by disrupting the KCNK3 gene in the T-47D human breast ductal carcinoma epithelial cell line. This polyclonal knockout pool provides a loss-of-function model for studying the TASK-1 potassium channel without clonal isolation.
The T-47D cell line was originally derived from the pleural effusion of a 54-year-old female with ductal carcinoma of the breast. These cells are estrogen receptor (ER)-positive, progesterone receptor (PR)-positive, and androgen receptor (AR)-positive, making them a well-established model for luminal A breast cancer. T-47D cells exhibit estrogen-responsive growth and are widely used to investigate hormone-dependent signaling pathways in breast cancer biology.
KCNK3 encodes the TASK-1 (TWIK-related acid-sensitive K+ channel 1) two-pore domain potassium channel that mediates background K+ currents, thereby setting the resting membrane potential and regulating cellular excitability. TASK-1 channels respond to diverse extracellular and intracellular signals, including extracellular pH changes, hypoxia via HIF-1??, Gq-coupled receptor activation by angiotensin II or acetylcholine, and phosphorylation by protein kinases such as PKA, PKC, and SGK1. Downstream, TASK-1 influences intracellular Ca2+ levels, activating AKT and MAPK/ERK signaling cascades that modulate cell cycle regulators like cyclin D1 and apoptotic factors such as Bcl-2 family members. TASK-1 also interacts with TASK-3 (KCNK9) channels, 14-3-3 proteins, ??-arrestin, and the coatomer protein complex, positioning KCNK3 at a signaling hub linking membrane potential to critical cellular processes.
Knockout of KCNK3 in T-47D breast cancer cells is predicted to depolarize the membrane potential, altering calcium influx and downstream signaling pathways, including the AKT and MAPK/ERK axes. Given that T-47D cells are hormone-sensitive and express ER, PR, and AR, KCNK3 disruption may influence hormone-driven proliferation, survival, and response to therapeutic agents. This polyclonal knockout model thus enables researchers to dissect the role of TASK-1 in breast cancer progression, hypoxia adaptation, and cross-talk between ion channels and steroid receptor signaling.
This knockout cell population is suitable for a broad range of experimental approaches, including western blotting and RT-qPCR to confirm target gene disruption, RNA-seq for transcriptomic profiling, patch clamp electrophysiology to assess K+ current changes, and functional assays such as MTT proliferation assays, Annexin V apoptosis assays, calcium imaging, and Transwell migration assays. The cells can be employed to validate KCNK3 as a therapeutic target in breast cancer and pulmonary arterial hypertension, to screen TASK-1 modulators, and to investigate hypoxia-induced signaling pathways. For further information, please contact Ascent Research.
