The EBAG9 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji human B lymphoblast cell line, featuring targeted disruption of the EBAG9 gene. This loss-of-function model enables investigation of EBAG9-dependent processes without defined clonal selection, providing a heterogeneous population that mirrors physiological genetic variability.
The Raji host cell line originates from an EBV-positive Burkitt’s lymphoma and retains characteristics of mature B lymphoblasts, including immunoglobulin production and antigen presentation functions. These cells serve as a well-established model for B-cell biology, lymphomagenesis, and adaptive immunity, offering a relevant background for studying immune regulatory genes in a transformed B-cell context.
EBAG9 encodes an estrogen-regulated type II transmembrane protein that functions as a ligand for a putative RCAS1 receptor on immune effector cells. Upon receptor engagement, EBAG9 triggers apoptosis through caspase activation, involving downstream effectors such as caspase-3, cytochrome c, and BAX. Expression of EBAG9 is upregulated by the estrogen receptor alpha (ESR1) in response to estradiol, linking hormonal signaling to immune evasion. This axis promotes depletion of tumor-infiltrating lymphocytes, contributing to the immune escape of estrogen-responsive malignancies.
In the Raji B lymphoblast background, EBAG9 knockout provides a unique tool to dissect the interplay between B-cell survival and immune cell cytotoxicity. Raji cells naturally express EBV latency genes that modulate apoptosis and immune recognition; disrupting EBAG9 in this context allows researchers to examine how loss of an immunomodulatory ligand affects crosstalk with T cells and NK cells. This model is particularly valuable for understanding EBAG9??s role in B-cell lymphomagenesis and for testing whether its inhibition can restore anti-tumor immunity.
Researchers can employ this polyclonal knockout population in a variety of experimental setups, including co-culture apoptosis assays monitored by flow cytometry to assess immune cell killing, cytotoxicity assays to quantify target cell death, and molecular analyses such as Western blotting, RT-qPCR, and RNA-seq to profile signaling changes. Applications span tumor immunology, immune checkpoint research, and development of cancer immunotherapies, with particular relevance to ovarian, endometrial, and prostate cancers where EBAG9-mediated immune evasion is documented. For additional information or custom requests, please contact Ascent Research.
