The KRT14 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population for disruption of the KRT14 gene in human gastric adenocarcinoma cells. The polyclonal pool contains genetically diverse cells with targeted inactivation of KRT14, avoiding clonal selection biases. This loss-of-function model facilitates studies of keratin 14 in an epithelial cancer context.
The AGS parental line is an adherent epithelial cell model derived from a human gastric adenocarcinoma, widely used for gastric cancer research. It retains epithelial characteristics, including expression of keratins, and provides a relevant platform to assess the impact of KRT14 knockout on tumor cell behavior.
KRT14 encodes a type I intermediate filament protein that obligately pairs with its type II partner KRT5 to form cytoskeletal networks anchored at desmosomes and hemidesmosomes through junctional proteins desmoplakin (DSP), junction plakoglobin (JUP), and plakophilin-1 (PKP1). Its transcription is controlled by upstream regulators TP63, AP-1 family transcription factors, and TGFB1, while downstream it influences cell adhesion molecules and AKT1 signaling. Disruption of KRT14 dismantles the KRT5?CKRT14 filament network and compromises desmosome integrity, thereby reducing epithelial mechanical stability and altering cell migration and signal transduction.
In the context of AGS gastric cancer cells, KRT14 knockout disrupts intermediate filament architecture, potentially weakening cell?Ccell adhesions and promoting a mesenchymal phenotype, or alternatively impairing migration if the cytoskeleton fails to support invasion. This directly impacts epithelial-mesenchymal transition-related processes and may alter cellular responses to anoikis and chemotherapeutics. The model is valuable for examining how keratin loss intersects with AKT1 signaling and contributes to gastric cancer progression.
This polyclonal knockout pool is suited for western blot and immunofluorescence analyses to verify KRT14 ablation and monitor compensatory changes in KRT5, DSP, or other desmosomal proteins. RT-qPCR can quantify EMT markers such as vimentin and E-cadherin. Functional assays including Boyden chamber migration, wound healing, and adhesion to extracellular matrix proteins directly test the role of KRT14 in motility and attachment. Flow cytometry permits profiling of epithelial surface markers. These tools enable detailed investigation of gastric cancer cell behavior. For further technical inquiries, please contact Ascent Research.
