The AKT3 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the AGS human gastric adenocarcinoma cell line. This polyclonal pool carries targeted disruption of the AKT3 gene, facilitating loss-of-function studies of the serine/threonine kinase AKT3. By eliminating AKT3 expression, the model enables dissection of AKT3-specific functions within the gastric cancer context, distinguishing it from the closely related AKT1 and AKT2 isoforms.
The AGS cell line, established from a patient with gastric adenocarcinoma, exhibits adherent epithelial morphology and is widely used as an in vitro model for gastric cancer research. These cells retain key signaling pathway alterations relevant to gastric carcinogenesis, making them an appropriate host for studying oncogenic mechanisms and therapeutic vulnerabilities. The AGS background provides a physiologically relevant system for investigating AKT3-mediated signaling in a disease-relevant cellular environment.
AKT3 is a critical effector of the PI3K/AKT signaling cascade. It is activated downstream of growth factor receptors such as EGFR and IGF-1R, through PI3K-generated PIP3-mediated membrane recruitment, where it is phosphorylated by PDK1 (Thr308) and mTORC2 (Ser473). Upon activation, AKT3 phosphorylates a diverse set of downstream targets, including GSK3??, FOXO transcription factors, mTORC1 (via PRAS40 and TSC2), MDM2, BAD, and Caspase-9, thereby promoting cell survival, proliferation, metabolism, and growth. Interacting factors such as PIP3, TSC1/TSC2 complex, and PRAS40 modulate its activity. This kinase thus integrates mitogenic and nutrient signals to coordinate key cellular processes.
In gastric adenocarcinoma, hyperactivation of the PI3K/AKT pathway, often through PTEN loss or PIK3CA mutations, drives tumor progression. AKT3, in particular, has been implicated in gastric cancer cell proliferation, resistance to apoptosis, and enhanced migratory capacity. The AKT3 knockout in AGS cells creates a powerful loss-of-function model to directly assess the contribution of AKT3 to these malignant phenotypes, independent of other AKT isoforms. This model is instrumental for studying AKT3-dependent signaling networks and evaluating isoform-specific therapeutic targeting.
Researchers can employ this polyclonal knockout cell population in a variety of functional assays to investigate PI3K/AKT pathway dynamics and gastric cancer biology. Typical applications include western blot analysis of phosphorylated AKT substrates (e.g., p-GSK3??, p-FOXO), cell viability assays (MTT or CellTiter-Glo), colony formation assays, Transwell migration and invasion studies, and apoptosis assays (Annexin V/PI staining). The polyclonal knockout format is particularly suited for studying population-level responses and pathway dependencies in gastric cancer. For detailed product information and technical support, please contact Ascent Research.
