The EBAG9 Knockout NCI-H1299 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt expression of the EBAG9 gene in the human non-small cell lung cancer cell line NCI-H1299. This loss-of-function model leverages CRISPR/Cas9-mediated gene disruption to generate a heterogeneous pool of cells, each carrying targeted modifications at the EBAG9 locus. The polyclonal format preserves the genetic diversity inherent to the edited population, enabling robust studies of gene ablation effects within a physiologically relevant cell context. This product serves as a powerful tool for investigating tumor immune evasion mechanisms and evaluating therapeutic strategies aimed at restoring antitumor immunity.
NCI-H1299 cells are derived from the lymph node metastasis of a lung adenocarcinoma, representing an aggressive, metastatic non-small cell lung cancer model. Widely employed in oncology research, this cell line exhibits rapid proliferation, anchorage-independent growth, and potent invasive capacity, mirroring key aspects of advanced lung cancer. Its tumorigenic properties and well-characterized signaling networks make NCI-H1299 an ideal host for studying genes involved in metastasis, immune escape, and tumor microenvironment interactions. The cells provide a clinically relevant background for assessing functional consequences of EBAG9 loss.
EBAG9 (RCAS1) is a tumor-associated antigen that acts as a potent inhibitor of antitumor immunity by triggering apoptosis in tumor-infiltrating lymphocytes. Mechanistically, EBAG9 binds to an undefined receptor on T cells and natural killer (NK) cells, leading to cytochrome c release, caspase activation, and programmed cell death. Expression of EBAG9 is regulated downstream of estrogen receptor alpha (ESR1) and estrogen signaling, linking hormonal pathways to immune suppression. Key pathway components include EBAG9/RCAS1, caspases, and cytochrome c, with downstream effects involving inactivation of immune effector cells and promotion of tumor progression through immune evasion and apoptosis signaling networks.
In the NCI-H1299 context, EBAG9 contributes to the cell line??s ability to evade immune surveillance, mirroring the immune escape mechanisms observed in lung cancer and other malignancies. Disruption of EBAG9 is predicted to abolish this immunosuppressive function, rendering tumor cells susceptible to immune-mediated clearance. This knockout model thus provides a unique platform to dissect tumor-immune cell crosstalk, particularly through co-culture experiments with peripheral blood mononuclear cells (PBMCs), where enhanced T cell and NK cell viability and increased tumor cell apoptosis can be quantified.
This cell model is suited for diverse research applications in cancer immunology, tumor microenvironment studies, immune checkpoint research, and drug discovery. Representative assays include co-culture apoptosis assays with PBMCs, flow cytometry to assess immune cell viability and caspase activation, western blotting for cytochrome c release and cleavage of caspases, migration and invasion assays, and drug sensitivity profiling. These applications enable detailed exploration of apoptosis signaling, immune resistance, and therapeutic vulnerabilities. For further information, please contact Ascent Research.
