The EFNB1 Knockout K-562 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of K-562 cells in which the EFNB1 gene has been disrupted to eliminate expression of the ephrin-B1 ligand. This product provides a heterogeneous knockout pool, enabling loss-of-function studies without clonal isolation, and serves as a versatile model for interrogating ephrin-B1-dependent signaling in a leukemic background.
The K-562 host cell line was derived from a 53-year-old female patient with BCR-ABL-positive chronic myelogenous leukemia in blast crisis. These lymphoblastoid cells are a widely established model for hematopoietic differentiation and leukemia biology, characterized by active BCR-ABL tyrosine kinase signaling, which drives proliferation and survival, and by the capacity to differentiate along erythroid, megakaryocytic, and myeloid lineages under appropriate stimuli.
Ephrin-B1, encoded by EFNB1, is a transmembrane ligand that engages Eph receptor tyrosine kinases, most notably EphB2 and EphA4, to initiate bidirectional signaling. Forward signaling downstream of Eph receptors mobilizes Src family kinases, focal adhesion kinase (FAK), and the MAPK/ERK cascade, whereas reverse signaling through the ephrin-B1 cytoplasmic tail recruits PDZ-domain adaptors such as GRIP1 and modulates Rho family GTPases (RhoA, Rac1). Ephrin-B1 expression is regulated by transcription factors including HIF-1??, NF-??B, and Notch, and its function is integrated with integrin-mediated adhesion, coupling cell?Ccell repulsion and attachment to cytoskeletal reorganization and motility.
In the K-562 context, disruption of EFNB1 is expected to uncouple Eph receptor forward and reverse signaling, potentially altering MAPK/ERK and PI3K/AKT pathway activity and disturbing Rho GTPase-driven cytoskeletal dynamics. This perturbation can affect leukemic cell adhesion, migration, and proliferation, offering a defined genetic tool to dissect the contribution of ephrin-B1 to CML pathophysiology and to evaluate the dependency of BCR-ABL-driven signaling on Eph-ephrin crosstalk.
Researchers can employ these polyclonal knockout cells to investigate ephrin-B1 function in cancer cell signaling, hematologic malignancy progression, and drug resistance. Representative applications include Transwell migration and invasion assays, Rho GTPase activation pull-downs, Western blotting for ephrin-B1 and phospho-Eph receptors, RT-qPCR for EFNB1 transcript levels, flow cytometric analysis of Eph receptor surface expression, co-immunoprecipitation of Eph-ephrin complexes, and in vitro drug sensitivity screens targeting Eph kinases or downstream effectors. For further technical specifications and ordering information, please contact Ascent Research.
