The CCDC91 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population for targeted disruption of the CCDC91 gene in AGS gastric adenocarcinoma epithelial cells. This heterogeneous pool of cells carries diverse loss-of-function mutations at the CCDC91 locus, enabling functional studies without the constraints of clonal selection. The knockout is achieved through CRISPR/Cas9-mediated gene disruption, generating a population that collectively represents a complete knockout background for downstream analyses.
The parental AGS cell line originates from a human gastric adenocarcinoma isolated from a 54-year-old female patient. These cells exhibit an adherent epithelial morphology and are widely used as a model system for gastric cancer research, including studies on proliferation, metastasis, and drug response. The AGS line retains characteristic signaling aberrations common to gastric adenocarcinomas, including dysregulated Wnt/??-catenin pathway activity, making it a highly relevant host for investigating CCDC91 function in a disease-appropriate cellular context.
CCDC91 positively regulates Wnt/??-catenin signaling by interacting with ??-catenin and preventing its phosphorylation by GSK-3?? within the destruction complex (AXIN1, APC). This protects ??-catenin from degradation, leading to its stabilization and nuclear translocation, where it partners with TCF/LEF to activate targets like CCND1, MYC, MMP7, and BIRC5. Upstream, WNT3A binding to Frizzled receptors and Dishevelled initiates the pathway, converging on ??-catenin. CCDC91 thus sustains Wnt transcriptional output in gastric cancer cells by stabilizing ??-catenin.
In the AGS gastric adenocarcinoma background, CCDC91 upregulation contributes to enhanced cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT), processes fundamental to tumor progression and metastasis. Disruption of CCDC91 in these polyclonal knockout cells abrogates ??-catenin stabilization, thereby attenuating Wnt target gene expression and reducing the aggressive phenotype, providing a physiologically relevant model to study the dependence of gastric cancer on aberrant Wnt signaling.
These polyclonal knockout cells enable applications such as western blotting for total and active ??-catenin, TOP/FOP flash luciferase reporter assays for TCF/LEF activity, RT-qPCR of Wnt target genes (CCND1, MYC, MMP7, BIRC5), cell proliferation (CCK-8), and migration/invasion assays. Additional uses include immunofluorescence for ??-catenin localization, co-immunoprecipitation to probe CCDC91?C??-catenin interactions, xenograft tumor models, and drug sensitivity assays. For detailed technical specifications and support, please contact Ascent Research.
