The EFNA5 Knockout HGC-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for the investigation of ephrin-A5 signaling in gastric adenocarcinoma. This product utilizes CRISPR/Cas9-mediated gene disruption to target EFNA5 in the HGC-27 human gastric carcinoma epithelial cell line, generating a heterogeneous pool of cells with disrupted EFNA5 expression. The polyclonal format offers a versatile tool for studying gene function without the clonal selection bias inherent in monoclonal lines, making it suitable for functional assays that probe collective cell behavior, such as tumor cell migration and invasion.
The HGC-27 cell line is an epithelial model derived from the lymph node metastasis of a patient with gastric adenocarcinoma. This aggressive, metastatic origin renders HGC-27 cells particularly valuable for studying advanced gastric cancer and mechanisms of metastasis. The cells exhibit characteristics of poorly differentiated adenocarcinoma and are widely employed in cancer biology research to investigate signaling pathways underlying tumor progression, cell adhesion, and invasive potential.
EFNA5 encodes ephrin-A5, a glycosylphosphatidylinositol (GPI)-anchored ligand for Eph receptor tyrosine kinases. Ephrin-A5 mediates bidirectional signaling through interactions with Eph receptors such as EphA4, EphA2, EphA3, and EphB2, as well as through reverse signaling into the ephrin-expressing cell. Upon receptor binding, ephrin-A5 triggers activation of Src and focal adhesion kinase (FAK), leading to regulation of RhoA and actin cytoskeleton dynamics. Downstream effectors include the RAS-MAPK/ERK1/2 cascade, which modulates cell proliferation and migration. Upstream, ephrin-A5 expression and shedding are controlled by TGF-?? signaling, the Wnt/??-catenin pathway, and ADAM10/17 sheddases. Integrins also serve as important interaction partners, contributing to the crosstalk between cell adhesion and Eph-ephrin signaling.
In the context of HGC-27 cells, EFNA5 knockout likely disrupts both forward and reverse Eph-ephrin signaling, impairing the coordination of cell adhesion and repulsion events critical for collective cell migration and invasion. Given the metastatic origin of the host cells, this polyclonal knockout pool provides a physiologically relevant model to dissect the contribution of ephrin-A5 to gastric cancer metastasis. The loss of EFNA5 may compromise the activation of PI3K/AKT and MAPK/ERK pathways, thereby attenuating the invasive properties of these cells. This model also permits the study of how ephrin-A5 modulates interactions within the tumor microenvironment.
Key applications include quantitative analysis of cell migration and invasion using Transwell or scratch-wound assays, adhesion assays on extracellular matrix substrates, and immunofluorescence staining to visualize actin cytoskeletal rearrangements. Researchers can validate EFNA5 disruption via Western blotting and assess downstream signaling changes through phosphorylation-specific antibodies for EphA4, Src, FAK, and ERK1/2. Transcriptomic profiling via RNA-seq can reveal broader gene expression alterations upon EFNA5 loss. Co-immunoprecipitation assays allow investigation of altered Eph receptor interactions, and drug sensitivity screens with Eph receptor inhibitors can evaluate therapeutic targeting. For detailed product inquiries, please contact Ascent Research.
