The ANO10 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. This product provides a loss-of-function model for the ANO10 gene, enabling detailed investigation of its roles in cellular signaling, ion homeostasis, and cancer biology. The polyclonal format preserves genetic diversity within the cell population, offering a robust and representative system for functional genomics studies without the clonal biases inherent in monoclonal lines.
The HT29 host cell line is a well-established human female colorectal adenocarcinoma model exhibiting epithelial morphology. Under appropriate conditions, HT29 cells can differentiate into enterocyte-like cells, recapitulating aspects of intestinal epithelial function. This line carries mutations in key oncogenes and tumor suppressors, including APC and TP53, making it a standard platform for colorectal cancer research. Its ability to form polarized monolayers and its endogenous ANO10 expression provide a physiologically relevant context for assessing gene function.
ANO10 encodes a dual-function protein that operates as a calcium-activated chloride channel and a phospholipid scramblase. Mechanistically, ANO10 engages the epidermal growth factor receptor (EGFR) through interactions with clathrin and the AP-2 adaptor complex, facilitating EGFR internalization and subsequent lysosomal degradation. This action negatively regulates the GRB2?CSOS?CRAS?CRAF?CMEK?CERK signaling cascade, thereby attenuating downstream cellular proliferation and migration. Upstream, ANO10 is activated by calcium influx, EGF, and cAMP-dependent pathways, while its disruption leads to sustained MAPK signaling, altered chloride ion flux, and enhanced cell motility.
In the HT29 colorectal cancer context, knockout of ANO10 removes a critical brake on EGFR-MAPK pathway activity, resulting in increased proliferative and migratory capacities. This model thus serves as a powerful tool to dissect the interplay between EGFR trafficking, calcium signaling, and oncogenic progression. Researchers can utilize this system to explore how ANO10-mediated receptor downregulation suppresses tumor-like behavior, and to investigate the molecular underpinnings of ANO10-related spinocerebellar ataxia type 10 within a cancer-relevant cellular environment.
Key experimental applications include Western blot analysis of EGFR and phospho-ERK, immunofluorescence localization studies, calcium imaging, chloride flux assays, wound healing and transwell migration assays, MTT proliferation assays, co-immunoprecipitation of EGFR complexes, RNA-seq transcriptomic profiling, and flow cytometry for cell cycle distribution. The ANO10 Knockout HT29 Polyclonal Cells are suited for research into spinocerebellar ataxia mechanisms, colorectal and gastric cancer signaling, ion channel biology, and drug screening targeting EGFR pathway components or calcium-activated chloride channels. For further information, please contact Ascent Research.
