The EBAG9 Knockout K-562 Polyclonal Cells provide a versatile loss-of-function model generated through CRISPR/Cas9-mediated gene disruption in the K-562 human chronic myelogenous leukemia (CML) cell line. This polyclonal knockout cell population enables robust investigation of EBAG9-dependent mechanisms without clonal selection artifacts, offering a representative cellular pool for functional studies.
The parental K-562 cell line originates from the pleural effusion of a 53-year-old female with CML in blast crisis. These suspension-adapted lymphoblasts are characterized by the BCR-ABL oncogene and a p53-null background, serving as a widely used model for hematopoietic stem cell biology, erythroleukemia, and CML blast crisis. Their genetic context provides a disease-relevant platform to examine oncogenic signaling and immune interactions.
EBAG9 is an estrogen-responsive gene encoding the tumor-associated antigen RCAS1. RCAS1 functions as a ligand that induces apoptosis in tumor-infiltrating lymphocytes, facilitating immune evasion in the tumor microenvironment. At the molecular level, RCAS1 interacts with estrogen receptor alpha (ER??) to modulate estrogen signaling, while downstream activation of caspase-3 and release of cytochrome c mediate programmed cell death in target lymphocytes. Additionally, NF-??B signaling integrates upstream stimuli, including estrogen, to regulate EBAG9 expression, thereby connecting hormonal and immune-regulatory pathways.
In the K-562 model, EBAG9 knockout disrupts the RCAS1-mediated apoptotic signal delivered to immune effectors, potentially restoring T-cell cytotoxicity against leukemic cells. Given the BCR-ABL-driven and p53-deficient milieu, this knockout system permits dissection of ER??-associated pathways and NF-??B crosstalk in leukemia progression, while also offering a tool to explore interactions with putative RCAS1 receptors on lymphocytes. The polyclonal nature preserves the heterogeneity inherent to the original line, avoiding artifacts from single-cell subcloning.
Typical research applications include studying cancer immunotherapy, tumor immune escape mechanisms, and estrogen signaling in leukemia. This product is suited for western blotting, RT-qPCR, immunofluorescence, and flow cytometry to assess apoptosis using Annexin V staining. Co-immunoprecipitation with ER?? enables protein interaction studies, while co-culture with T cells and cytokine release assays permit functional evaluation of immune evasion. BCR-ABL signaling analysis can further elucidate the interplay between oncogenic drivers and immune-modulatory genes. For more detailed information, please contact Ascent Research.
