EBAG9 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human gastric adenocarcinoma AGS cell line, engineered to disrupt the EBAG9 gene. The polyclonal pool contains a heterogeneous mixture of cells with loss-of-function mutations in the target locus, providing a genetically diverse knockout model that avoids clonal artifacts. This product enables robust functional studies of EBAG9 without the need for single-cell clone isolation, making it suitable for pooled screening approaches and bulk analyses of tumor?Cimmune interactions.
AGS cells originate from a female patient with gastric adenocarcinoma and are widely employed as a model for gastric epithelial biology, including mucosal barrier function, secretion, and H. pylori infection research. The cell line retains key characteristics of gastric epithelium and is responsive to hormonal and inflammatory stimuli, establishing a relevant context for investigating estrogen signaling and immune evasion mechanisms in gastric carcinogenesis.
EBAG9, also known as RCAS1 (receptor-binding cancer antigen expressed on SiSo cells), encodes a tumor-associated antigen that functions as a ligand for a putative receptor, RCAS1-R, and possibly Siglec-9, on immune effector cells. Expression of EBAG9 is transcriptionally activated by estrogen receptor alpha (ESR1) and upregulated by tumor necrosis factor alpha (TNF-??). Upon binding to its receptor, EBAG9 induces apoptosis in T lymphocytes and natural killer (NK) cells through caspase-3 activation, facilitating immune escape. Additionally, EBAG9 interacts with Alix and the exosome marker CD63 to promote exosome biogenesis and secretion, thereby modulating the tumor microenvironment. Downstream, EBAG9 influences the PI3K/AKT signaling axis, further supporting tumor progression.
In the AGS gastric cancer context, EBAG9 knockout disrupts a key immune evasion pathway, allowing researchers to dissect the molecular mechanisms by which gastric cancer cells escape immune surveillance. The polyclonal knockout cells provide a model to examine how loss of EBAG9 affects apoptosis induction in co-cultured immune cells, exosome-mediated communication, and estrogen-driven tumor growth. This system is especially valuable for evaluating therapeutic strategies that target the RCAS1?CRCAS1-R interaction and for studying the interplay between hormonal signaling and immune checkpoint modulation.
Typical applications include Western blotting and RT-qPCR to confirm EBAG9 depletion, Annexin V-based apoptosis assays on immune cells co-cultured with the knockout cells, flow cytometry for apoptosis markers, immunofluorescence to assess EBAG9 localization, exosome isolation and characterization, co-immunoprecipitation to map interacting partners, and reporter assays for estrogen response elements. This knockout model supports investigations into gastric cancer immune evasion, tumor microenvironment dynamics, estrogen signaling in carcinogenesis, and exosome-mediated progression. For further information, contact Ascent Research.
