The EFNA1 Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the EFNA1 gene in the K-562 human chronic myelogenous leukemia (CML) cell line. This product provides a loss-of-function model for studying ephrin-A1 biological functions without introducing monoclonal artifacts. The polyclonal format retains the inherent heterogeneity of a non-clonal cell pool, enabling robust analysis of ephrin-A1-dependent processes in hematopoietic cells, including signal transduction, cell migration, and angiogenesis.
The K-562 host cell line, established from a 53-year-old female CML patient in blast crisis, is BCR-ABL1 positive and widely used as a model for hematopoietic differentiation and erythroleukemia. K-562 cells exhibit a blast-like phenotype and can be induced to differentiate along erythroid, granulocytic, and monocytic lineages. Their suspension growth and well-characterized signaling networks make them a convenient system for studying kinase-dependent pathways, integrin-mediated adhesion, and cytokine responses. The BCR-ABL1 oncoprotein constitutively activates multiple downstream effectors, including PI3K/AKT and MAPK/ERK, providing a disease-relevant context to examine how ephrin-A1 signaling intersects with leukemogenic pathways.
EFNA1 encodes ephrin-A1, a glycosylphosphatidylinositol (GPI)-anchored ligand that binds EphA receptor tyrosine kinases, triggering bidirectional signaling. Reverse signaling through ephrin-A1 engages Src family kinases and Rho family GTPases such as RhoA and Rac1, modulating cytoskeletal dynamics. Forward signaling via EphA2, the predominant EphA receptor in many cell types, recruits and phosphorylates Src and focal adhesion kinase (FAK), leading to activation of PI3K/AKT and MAPK/ERK cascades. Upstream regulators include hypoxia-inducible factor 1-alpha (HIF-1??) and ERK1/2, and ephrin-A1 interacts with EphA1?CEphA8, integrins, and ADAM10, which cleaves ephrin-A1 to terminate signaling.
In the K-562 model, EFNA1 knockout disrupts ephrin-A1-mediated signaling, allowing dissection of its contributions to hematopoietic cell behavior. Since K-562 cells express EphA receptors and depend on integrin-mediated adhesion for certain functions, loss of ephrin-A1 can alter cell-cell and cell-matrix interactions. The BCR-ABL1-driven oncogenic background shares common downstream pathways such as PI3K/AKT and MAPK/ERK with ephrin-A1, making this knockout valuable for delineating ephrin-A1-specific effects in leukemia. The polyclonal knockout format avoids clonal selection bias, enabling assessment of heterogeneous responses in adhesion, migration, and differentiation.
This knockout cell product is suited for functional assays including Western blotting for EphA2 and phospho-AKT/ERK, RT-qPCR for EFNA1, flow cytometry for ephrin-A1, Boyden chamber migration/invasion assays, co-immunoprecipitation of ephrin-A1/EphA2, and phospho-signaling arrays. Apoptosis assays can probe survival functions. These cells support cancer biology, angiogenesis, cell signaling, and tumor microenvironment studies. For further details, please contact Ascent Research.
