The ATP1B1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population generated from the HT29 human colorectal adenocarcinoma line, designed for targeted disruption of the ATP1B1 gene. This loss-of-function model enables the study of ATP1B1-dependent processes in a genetically heterogeneous pool, offering a representative system for functional analyses without artifacts from monoclonal selection.
The HT29 parental line is a widely utilized epithelial model derived from a human colorectal adenocarcinoma. Known for its mucin production and capacity to form polarized monolayers, HT29 provides a relevant background for investigating ion homeostasis, cell adhesion, and signaling pathways in intestinal epithelial biology and colorectal cancer.
ATP1B1 encodes the ??-subunit of Na?/K?-ATPase, which complexes with the ??-subunit ATP1A1 to maintain transmembrane ion gradients. The ??-subunit interacts with ankyrin and caveolin-1, anchoring the pump and regulating SRC kinase activity. Expression is driven by Sp1 and modulated by thyroid hormone, corticosteroids, and osmotic stress. Knockout disrupts SRC phosphorylation, altering ERK1/2 and CREB signaling, and intersects with the PI3K/AKT pathway through EGFR and MAPK1, thereby affecting cell cycle regulators, adhesion, and migration.
In HT29 colorectal cancer cells, ATP1B1 loss perturbs ionic balance and SRC-mediated cell adhesion, providing a model to study mechanisms of cancer progression and metastasis. The mucin-producing capacity of HT29 may also be impacted, linking ATP1B1 to epithelial barrier disruption and signal transduction alterations critical for invasion.
This polyclonal knockout cell population supports diverse assays, including ion flux measurements, cell migration and invasion analyses, and drug sensitivity screening. Molecular characterization via western blotting, RT-qPCR, and co-immunoprecipitation can elucidate changes in the SRC/ERK1/2/CREB network, while immunofluorescence and SRC phosphorylation assays probe adhesion complexes and kinase activity. RNA-seq enables transcriptome-wide insights into pathway rewiring. For technical details, please contact Ascent Research.
