The EBAG9 Knockout 786-O Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal cell population originating from the human 786-O renal cell carcinoma line. This product achieves targeted disruption of the EBAG9 gene, which encodes the estrogen-responsive tumor-associated antigen RCAS1. By utilizing a polyclonal format without single-cell cloning, these cells maintain genetic heterogeneity and provide a robust loss-of-function model. This approach enables investigation of EBAG9??s roles in cancer biology and immune modulation without claims of monoclonality or complete gene ablation.
The 786-O cell line is a widely employed human clear cell renal cell carcinoma (ccRCC) model, derived from a primary tumor. These adherent epithelial cells are characterized by VHL inactivation, leading to constitutive activation of hypoxia-inducible pathways. This genetic background makes them particularly suitable for dissecting mechanisms of kidney cancer progression, metastasis, and drug resistance. Moreover, their immunogenic nature and responsiveness to checkpoint inhibitors render them valuable for tumor?Cimmune interaction studies.
EBAG9, also known as RCAS1, is an estrogen-inducible type II transmembrane protein that functions as a ligand for a putative receptor on T lymphocytes and natural killer (NK) cells. Receptor engagement triggers apoptosis in these immune effectors through caspase activation and cytochrome c release, facilitating immune evasion. EBAG9 expression is transcriptionally regulated by estrogen-bound estrogen receptor alpha (ER??) and the transcription factor SP1. The downstream apoptotic signaling involves Bcl-2 family members, positioning EBAG9 at a critical node connecting hormonal signals to adaptive immune suppression.
In the context of 786-O ccRCC cells, disruption of EBAG9 impairs the tumor??s ability to induce immune cell apoptosis, potentially restoring anti-tumor immunity. Renal cell carcinomas often exploit RCAS1-mediated killing to evade infiltrating lymphocytes, contributing to therapy resistance. This knockout model thus permits detailed dissection of estrogen-dependent and -independent immune evasion mechanisms. Co-culture experiments with T and NK cells can evaluate changes in cytotoxicity, cytokine secretion, and the impact of estrogen stimulation on immune escape.
Typical applications include cancer immunotherapy, tumor immune evasion, and estrogen receptor signaling research. Compatible assays range from Western blotting and RT-qPCR for expression analysis to flow cytometry and Annexin V staining for apoptosis quantification. Co-culture immune killing assays and estrogen stimulation experiments further enable functional studies. These tools accelerate EBAG9 target validation and therapeutic screening. For additional details, please contact Ascent Research.
