The FPGT Knockout AGS Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human gastric adenocarcinoma cell line AGS. This product provides a loss-of-function model for the FPGT gene, which encodes fucose-1-phosphate guanylyltransferase, the enzyme responsible for the synthesis of GDP-fucose??the essential donor substrate for all fucosylation reactions. The polyclonal format offers a pooled population of edited cells, enabling robust and reproducible investigation of fucosylation-dependent processes without the clonal selection artifacts that may arise from single-cell-derived lines. Researchers can employ this model system to dissect how global fucosylation influences epithelial cell biology in a gastric cancer background.
The AGS host cell line is a well-characterized model of human gastric mucosal epithelial cells derived from a gastric adenocarcinoma. These adherent epithelial cells retain key features of gastric epithelium and are widely used to study gastric cancer pathogenesis, including proliferation, migration, and signaling aberrations. The AGS background provides a relevant cellular context for probing the role of fucosylation in gastric malignancy, as aberrant glycosylation patterns are hallmarks of cancer progression. The knockout of FPGT in this cell line allows for direct interrogation of fucose metabolism and its impact on oncogenic signaling networks in a disease-relevant microenvironment.
FPGT catalyzes the conversion of fucose-1-phosphate and GTP to GDP-fucose, a reaction that sits at the nexus of fucose salvage and de novo synthesis. GDP-fucose is indispensable for the activities of fucosyltransferases (FUT1?CFUT11) and protein O-fucosyltransferases (POFUT1, POFUT2), which modify glycoproteins and glycolipids. Consequently, FPGT functions upstream of critical signaling pathways; its disruption leads to a loss of fucosylation on Notch receptors and EGFR, both of which rely on O-fucose modifications for proper trafficking and ligand-induced activation. Upstream regulators such as HIF1A and TGFB1 modulate FPGT expression, linking fucosylation capacity to hypoxia and growth factor cues. The FPGT product GDP-fucose also serves as a substrate for GMDS-dependent pathways, and FPGT interacts with fucose-1-phosphate kinase to maintain nucleotide sugar pools. Thus, FPGT knockout ablates a central node in glycoconjugate biosynthesis, with pleiotropic effects on cellular signaling.
In AGS gastric cancer cells, FPGT knockout eliminates GDP-fucose production, leading to global hypofucosylation. This defect impairs fucosylation-dependent Notch receptor activation, as POFUT1-mediated O-fucosylation is requisite for Notch?Cligand interaction and subsequent proteolytic cleavage by ADAM17. Similarly, EGFR fucosylation modulates receptor dimerization and downstream phosphorylation. Consequently, FPGT-edited AGS cells exhibit attenuated Notch and EGFR signaling, resulting in reduced proliferation, migration, and invasion??phenotypes that can be rescued by exogenous fucose supplementation. This model thus recapitulates the functional consequences of fucosylation deficiency in an epithelial malignancy context, making it a powerful tool for dissecting glycosylation-dependent malignant traits.
This knockout cell product is ideally suited for a range of investigative applications. Researchers can utilize lectin blotting with Aleuria aurantia lectin (AAL) to assess global fucose levels, perform flow cytometry for cell-surface fucosylated epitopes, and conduct Western blotting for cleaved Notch1 (NICD) and phosphorylated EGFR. Functional assays such as transwell migration/invasion and MTT proliferation assays quantify phenotypic changes, while RNA-seq analysis reveals compensatory changes in fucosyltransferase expression. Apoptosis can be monitored via caspase-3 activation assays. These tools enable detailed studies of fucosylation in gastric cancer progression, Notch signaling dynamics, metastasis, drug resistance, and glycocalyx remodeling. For further technical information, researchers are encouraged to contact Ascent Research.
