The KCNK3 Knockout HT29 Polyclonal Cells constitute a CRISPR/Cas9-mediated gene-edited polyclonal population of HT29 cells harboring disruption of the endogenous KCNK3 locus. This product provides a heterogeneous knockout pool that eliminates TASK-1 (K2P3.1) potassium channel expression without single-cell cloning, facilitating loss-of-function investigations into background potassium conductance in colorectal cancer models.
HT29 cells are a well-characterized human colorectal adenocarcinoma line derived from a female patient. They carry a gain-of-function missense mutation in TP53 (p.R273H) and a loss-of-function mutation in APC, while remaining microsatellite stable. As an intestinal epithelial model, HT29 is extensively used to study colorectal cancer signaling, drug resistance, and epithelial barrier function.
The KCNK3 gene product, TASK-1, is a two-pore-domain potassium channel that generates pH- and hypoxia-sensitive leak currents, setting the resting membrane potential. Upstream regulators include Gq-coupled receptors (AT1R, ETAR), protein kinase C, and diacylglycerol, while downstream signaling involves membrane depolarization, activation of voltage-gated calcium channels (Cav1.2), and calcineurin/NFAT-mediated transcription. TASK-1 also participates in protein complexes with 14-3-3, forms heterodimers with TASK-3, and is subject to regulatory interactions with SUMO proteins and ??-arrestin-2.
In HT29 colorectal cancer cells, loss of KCNK3 depolarizes the plasma membrane, altering calcium influx through Cav1.2 channels and thereby perturbing intracellular calcium dynamics. This disruption impacts calcium-dependent processes such as cell proliferation, apoptosis, and migration, processes already dysregulated by the p53 and APC mutations present in this line. Consequently, the KCNK3 knockout model enables dissection of how TASK-1-mediated electrical signaling integrates with oncogenic pathways to modulate tumor cell behavior.
These polyclonal knockout cells are suitable for diverse functional assays, including MTT proliferation, Annexin V apoptosis, scratch wound healing, and transwell invasion assays to examine colorectal cancer cell aggressiveness. Ion channel pharmacology can be investigated via patch-clamp electrophysiology, FLIPR membrane potential measurements, and Fluo-4 calcium imaging. Transcriptomic responses to KCNK3 disruption may be analyzed by RNA-seq and RT-qPCR. Additionally, the model supports drug sensitivity screening and can serve as a platform for studying TASK-1-related pulmonary arterial hypertension. For further information, contact Ascent Research.
