The EEA1 Knockout HGC-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the HGC-27 human gastric cancer epithelial cell line, featuring targeted disruption of the EEA1 gene. This polyclonal knockout product provides a heterogeneous mixture of cells carrying various gene-disruption events, enabling robust loss-of-function studies in an endogenously relevant genetic background.
HGC-27 is a widely characterized human gastric undifferentiated carcinoma cell line exhibiting epithelial morphology. Originally established from a metastatic site, this cell line retains hallmark features of aggressive gastric cancer, including rapid proliferation and perturbed signaling networks. Its use in endocytosis research is well established, making it an ideal host for interrogating the role of EEA1 in endosomal trafficking and receptor-mediated signaling pathways.
EEA1 encodes an early endosome autoantigen protein that functions as a critical tethering factor, facilitating homotypic fusion of early endosomes. Mechanistically, EEA1 bridges Rab5:GTP and phosphatidylinositol 3-phosphate (PI3P) on opposing membranes, a process essential for endosome docking and fusion. EEA1 is activated by Rab5 GTPase and interacts with Rabaptin-5, Rabex-5, and calmodulin. It acts downstream of PI3K/Vps34-mediated PI3P production and is regulated by growth factors such as EGF and HGF. EEA1 recruits endosomal SNAREs including Syntaxin 6, Syntaxin 13, and VAMP8, and intersects with the ESCRT machinery. Through these interactions, EEA1 orchestrates endosomal maturation and the sorting of internalized receptors, including EGFR, toward lysosomal degradation.
In HGC-27 gastric carcinoma cells, EEA1-directed early endosomal fusion is intimately linked to the attenuation of oncogenic signaling. Aberrant receptor trafficking, particularly of EGFR and other RTKs, is a hallmark of gastric cancer progression and metastasis. Disruption of EEA1 is predicted to impair endosome fusion, leading to altered spatial and temporal dynamics of receptor internalization and enhanced or prolonged signaling. This polyclonal knockout model enables investigation of how defective early endosome function contributes to gastric cancer pathogenesis, including sustained proliferative signaling, evasion of apoptosis, and metastatic potential.
This product is suited for a broad range of experimental applications, including endocytosis and membrane trafficking studies, cancer signaling and receptor degradation analysis, drug delivery and nanoparticle uptake mechanisms, and autophagy-endosome-lysosome pathway investigations. Representative assays include Western blotting for EEA1 and endosomal markers, immunofluorescence co-localization of EEA1 with transferrin, EGFR internalization and degradation kinetics, transferrin uptake assays, Rab5 activity pull-downs, co-immunoprecipitation of EEA1-Rab5 complexes, and phospho-RTK signaling arrays. High-content screening for trafficking modulators and live-cell imaging of endosome dynamics also benefit from this model. For additional information, please contact Ascent Research.
