This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T human embryonic kidney cell line, featuring targeted disruption of the EFNB2 gene. The editing strategy results in loss of ephrin-B2 protein expression, providing a stable loss-of-function model for studying ephrin-B2-dependent cellular processes. The polyclonal nature preserves the genetic heterogeneity introduced during editing, allowing functional studies without clonal isolation artifacts.
HEK293T cells are a widely used HEK293 derivative that constitutively expresses SV40 large T antigen, enhancing episomal plasmid replication. These adherent epithelial cells are renowned for their high transfectability and are a preferred host for protein production, lentivirus packaging, and transient expression studies. Their robust growth and well-mapped signaling pathways provide an ideal platform for functional investigation of EFNB2 knockout.
EFNB2 encodes ephrin-B2, a single-pass transmembrane ligand that engages EphB receptor tyrosine kinases (EphB1, EphB2, EphB3, EphB4) to initiate juxtacrine bidirectional signaling. Forward signaling through EphB receptors activates SRC family kinases and modulates cytoskeletal dynamics via RHOA and RAC1, while reverse signaling through ephrin-B2 triggers PDZ-dependent recruitment of adaptor proteins such as GRIP1, PICK1, and syntenin, along with SRC-mediated phosphorylation. ephrin-B2 function is regulated by upstream transcription factors including HIF1??, VEGFA, NOTCH1 intracellular domain, ETS1, and ??-catenin/TCF complexes. Downstream, ephrin-B2 controls ERK1/2 and AKT phosphorylation, RHOA activation, FAK phosphorylation, and VEGFR2 internalization, linking Eph?Cephrin signals to MAPK/ERK, PI3K/AKT, and Rho GTPase pathways. Furthermore, ephrin-B2 interacts directly with VEGFR2, ADAM10, and EphB receptors, positioning it at the intersection of angiogenic and cell migration networks.
The HEK293T background offers an excellent model for dissecting ephrin-B2 function because these cells express endogenous EphB receptors and relevant downstream effectors but do not require ephrin-B2 for viability. Knockout of EFNB2 disrupts bidirectional Eph?Cephrin signaling, allowing clear assessment of ephrin-B2-dependent effects on adhesion, repulsion, and migration. High transfectability enables straightforward rescue and structure-function experiments with wild-type or mutant ephrin-B2 constructs. Moreover, the polyclonal nature avoids clonal selection artifacts, providing a more representative view of ephrin-B2 biology.
This knockout cell population is a versatile tool for investigating ephrin-B2 in cancer cell migration and invasion, tumor angiogenesis, and neurodevelopmental disorders. Typical applications include wound healing and Transwell migration/invasion assays to quantify contact-dependent repulsion and motility, tube formation assays to model angiogenic processes, and phospho-signaling analysis (p-ERK, p-AKT) to interrogate downstream pathway activation. Co-immunoprecipitation studies can further probe EphB4 binding and PDZ protein interactions, while western blotting and immunofluorescence confirm knockout efficiency and protein localization. Researchers studying VEGFR2 co-activation, atherosclerosis, diabetic retinopathy, or Eph?Cephrin reverse signaling mechanisms will find this model especially valuable. For further technical details or assistance, please contact Ascent Research.
