The ATP4A Knockout HCT 116 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HCT 116 colorectal carcinoma cell line, engineered for loss-of-function studies of the ATP4A gene (Homo sapiens). This product comprises a heterogeneous pool of edited cells with targeted disruption of ATP4A, providing a physiologically relevant model to investigate the gastric H+/K+ ATPase alpha subunit in a non-gastric cancer context without clonal selection artifacts.
The parental HCT 116 line is an epithelial colorectal adenocarcinoma model with KRAS (G13D) and PIK3CA (H1047R) mutations, widely used in tumor biology and drug screening. Its adherent growth and genetic stability support functional genomics, signaling assays, and pharmacological studies. The knockout pool maintains this tumorigenic background, enabling study of ATP4A in a colorectal cancer context.
ATP4A encodes the catalytic alpha subunit of the gastric H+/K+ ATPase, a pump that exchanges luminal K+ for cytoplasmic H+ to drive gastric acidification. Its activity is stimulated by histamine (H2 receptor), gastrin (CCK2 receptor), and acetylcholine (M3 receptor) via cAMP/PKA and CaMKII pathways, while somatostatin provides inhibitory input. The functional heterodimer with ATP4B depends on the KCNQ1/KCNE2 potassium channel for recycling, and downstream effects include proton secretion, pepsinogen activation, and CLIC6 chloride channel opening. Omeprazole and pantoprazole are direct inhibitors; ezrin anchors the complex to the cytoskeleton, and cross-regulation with EGF receptor ligands is documented.
Although normally gastric, ATP4A is ectopically expressed in some colorectal cancers. Knockout in HCT 116 allows study of its role in tumor pH regulation, ion flux, and signaling that intersect with oncogenic KRAS and PIK3CA. The polyclonal pool avoids clonal artifacts, providing a robust loss-of-function model. Researchers can assess effects on intracellular pH, potassium handling, and drug sensitivity to proton pump inhibitors, potentially exposing therapeutic targets.
These polyclonal knockout cells are ideally suited for a spectrum of investigative assays, including Western blotting and immunofluorescence to verify ATP4A loss and subcellular mislocalization, ATPase activity measurements, and pH-sensitive fluorescent dye experiments to quantify proton transport defects. Technical applications extend to potassium uptake assays, RNA-seq transcriptomic profiling to uncover compensatory pathways, and drug sensitivity screens employing omeprazole or pantoprazole. The model supports CRISPR screening for novel regulators of ion homeostasis and provides a valuable tool for validating ATP4A as a therapeutic target in non-gastric malignancies. For technical inquiries or custom knockout cell services, please contact Ascent Research.
