The ATP1A3 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma cell line. This product offers a mixed population of cells with targeted disruption of the ATP1A3 gene, encoding the ??3 subunit of the Na+/K+-ATPase. The polyclonal format provides a heterogeneous loss-of-function model suitable for studying the collective impact of ATP1A3 ablation in an epithelial cancer background. By eliminating the Na+/K+-ATPase ??3 subunit, the cells enable investigation of disrupted active ion transport and its downstream consequences without the need for clonal selection, preserving population-level diversity that reflects the complexity of tumor biology.
The HT29 host cell line originates from a 44-year-old Caucasian female with colorectal adenocarcinoma. These epithelial cells differentiate into polarized monolayers, serving as a standard model for intestinal barrier studies and cancer research. They retain oncogenic signaling and metabolic alterations, enabling investigation of tumor biology, drug responses, and epithelial transport.
ATP1A3 encodes the ??3 subunit of the Na+/K+-ATPase, which couples ATP hydrolysis to active Na+ export and K+ import, sustaining electrochemical gradients. The ??3 subunit is regulated by transcription factors (CREB, NRF2), kinases (PKA, PKC), and hormonal signals (thyroid hormone, insulin), while the cardiac glycoside ouabain acts as a potent inhibitor. It interacts with auxiliary ATP1B subunits, FXYD proteins, caveolin-1, ankyrin, and spectrin, and scaffolds c-Src. Through these complexes, it modulates downstream SRC kinase, ERK1/2, and AKT, linking the MAPK/ERK and PI3K/AKT/mTOR pathways. The pump also indirectly controls NCX and NHE activity, thereby influencing intracellular Ca2+ dynamics.
In HT29 cells, ATP1A3 knockout disrupts active ion transport, altering membrane potential, ion gradients, and cell volume. The loss of ??3 impairs Src-mediated activation of ERK and AKT pathways, which are critical for proliferation, survival, and migration. This polyclonal knockout model thus enables dissection of how ATP1A3 contributes to colorectal cancer pathophysiology, including potential roles in drug resistance, epithelial-mesenchymal transition, and barrier integrity. It also facilitates exploration of crosstalk between ion homeostasis and oncogenic signaling in a non-neuronal context.
Typical applications include Na+/K+-ATPase activity assays, intracellular sodium/potassium flux measurements with SBFI/PBFI, calcium imaging (Fluo-4), proliferation (MTT), migration (wound healing), and protein analysis (western blot, RT-qPCR, immunofluorescence). Patch-clamp electrophysiology can assess membrane potential changes. This polyclonal knockout population provides a versatile tool for studying ATP1A3-related signaling in colorectal cancer. For further information, contact Ascent Research.
