The EBAG9 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HEK293T human embryonic kidney cells, generated to disrupt the EBAG9 gene. This gene encodes the receptor-binding cancer antigen (RCAS1), a tumor-associated factor implicated in immune evasion. The polyclonal nature provides a functionally heterogeneous model for studying EBAG9 loss-of-function without clonal selection artifacts.
HEK293T cells are an immortalized adherent line expressing the SV40 large T antigen, which promotes episomal plasmid replication and high-level protein expression. Widely applied in molecular biology and virology, these cells offer a robust and well-characterized platform for gene-editing applications. Their ease of transfection and adaptability to diverse experimental formats make them an ideal host for investigating the molecular consequences of EBAG9 knockout in a controlled cellular environment.
EBAG9 expression is regulated by estrogen receptors alpha and beta, linking its activity to hormone-dependent cancers. The protein product, RCAS1, is a membrane-associated ligand that binds to putative receptors on T lymphocytes and natural killer cells, triggering caspase-3 and caspase-9 activation and subsequent apoptosis. This receptor-mediated killing inhibits anti-tumor immune responses, contributing to the progression of malignancies including breast, prostate, uterine, ovarian, and lung cancers.
In HEK293T cells, CRISPR/Cas9-mediated disruption of EBAG9 abolishes RCAS1 production, eliminating the capacity to induce apoptosis in immune effector cells. This knockout model enables straightforward dissection of the RCAS1 signaling axis through co-culture assays and permits complementation studies by re-introducing EBAG9 variants. The HEK293T background also supports efficient viral vector production for secondary gene delivery, facilitating investigation of pathway components in parallel loss- and gain-of-function experiments.
Primary applications encompass immune evasion research, apoptosis signaling analysis, and cancer immunology. Co-culture with T cells or NK cells allows direct measurement of lymphocyte apoptosis and activation status. The polyclonal cells serve as negative controls for antibody validation and as a starting material for further clonal isolation. They are also suitable for protein?Cprotein interaction studies and functional genomic screens. For additional information, please contact Ascent Research.
