The ATXN3 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. This heterogeneous pool carries a targeted disruption of the ATXN3 gene, resulting in a loss-of-function model. The polyclonal format reduces clonal variation artifacts and is well suited for pooled functional genomics screens and bulk biochemical analyses.
HT29 cells are an established epithelial model of human colorectal adenocarcinoma, capable of differentiation into enterocyte-like cells under defined conditions. This feature makes them invaluable for investigating intestinal epithelial biology, colorectal cancer pathogenesis, and gut barrier function. Their widespread use in drug discovery and signaling studies provides a robust context for functional gene analysis.
ATXN3 is a deubiquitinating enzyme that cleaves polyubiquitin chains, thereby regulating protein turnover through the ubiquitin-proteasome system. It participates in ER-associated degradation, aggrephagy, and the DNA damage response. Upstream regulators include NRF2 and HSF1, while MAPK and FOXO signaling pathways control its activity. ATXN3 interacts directly with VCP/p97, HSP70/HSC70, and HDAC6, forming complexes that influence substrate deubiquitination. Downstream, ATXN3 modulates p53 stability and activity, as well as CHIP/STUB1, Parkin, and PTEN. By deubiquitinating p53 and other targets, ATXN3 affects cell cycle control, apoptosis, and proteasomal degradation, thereby integrating stress signals with protein quality control.
In the HT29 colorectal cancer background, ATXN3 knockout provides a model to dissect its contributions to tumor cell biology. ATXN3??s regulation of p53 is particularly relevant, as p53 mutations are common in colorectal cancer and influence therapeutic responses. Additionally, ATXN3??s role in clearing misfolded proteins via aggrephagy and ERAD is critical for cancer cells experiencing proteotoxic stress from rapid proliferation or chemotherapeutics. Disrupting ATXN3 can reveal dependencies that may be exploited for targeted interventions.
Typical applications include deubiquitinase activity profiling, ubiquitination assays, and protein aggregation/clearance studies. The cells are suitable for deubiquitinase inhibitor screening, co-immunoprecipitation to assess protein?Cprotein interactions, and immunofluorescence microscopy. Western blotting and RT-qPCR enable quantification of expression changes in ATXN3 and its effectors. Additionally, the polyclonal knockout population facilitates high-throughput phenotypic screens to identify modifiers of ATXN3-associated pathways. For further details, please contact Ascent Research.
