The EFNB3 Knockout HEK293T Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population derived from HEK293T cells, in which the EFNB3 gene has been disrupted to eliminate expression of the ephrin-B3 protein. This heterogeneous polyclonal pool provides a versatile loss-of-function model for dissecting the molecular functions of ephrin-B3, avoiding the selection bias associated with monoclonal knockout lines and better reflecting population-level genetic diversity.
The parental HEK293T cell line is a human embryonic kidney epithelial cell line immortalized via stable integration of the SV40 large T antigen and adenovirus 5 E1A/E1B genes. Renowned for its high transfection efficiency and capacity for robust recombinant protein production, HEK293T is a widely used platform for studying signal transduction cascades, protein?Cprotein interactions, and functional genomics.
Ephrin-B3, encoded by EFNB3, is a GPI-anchored ligand for Eph receptor tyrosine kinases, engaging principally EphA4, EphB2, and EphB4 in bidirectional signaling. Forward signaling via Eph receptors and reverse signaling through the ephrin-B3 cytoplasmic domain recruit PDZ domain-containing adaptors such as GRIP1 and PICK1. Upstream, EFNB3 expression is transcriptionally regulated by retinoic acid, fibroblast growth factors, and homeobox transcription factors HOXA and PAX6. Upon activation, ephrin-B3 modulates small Rho GTPases (RhoA, Rac1, CDC42), Src family kinases, focal adhesion kinase (FAK), and downstream kinases ERK1/2 and AKT, as well as GSK-3??, thereby orchestrating cytoskeletal remodeling, cell repulsion, adhesion, and migration.
In the HEK293T background, which endogenously expresses multiple Eph receptors, EFNB3 knockout disrupts both reverse signaling and the ligand presentation necessary for forward signaling, creating a clean system to delineate ephrin-B3-specific contributions. This knockout pool eliminates confounding ligand-dependent Eph activation, enabling precise interrogation of ephrin-B3-mediated functions such as repulsive guidance cues and adhesion dynamics??processes critical in neurodevelopment, cancer metastasis, and cardiovascular morphogenesis. The polyclonal format preserves cellular heterogeneity, yielding more physiologically relevant data in population-based assays.
This product is optimized for a spectrum of research applications, including functional analysis of Eph-ephrin signaling, high-content screening for migration modulators, protein interaction mapping, and drug target validation. Compatible assays range from standard western blotting, RT-qPCR, immunofluorescence, and flow cytometry to advanced co-immunoprecipitation, transwell migration and invasion assays, phospho-kinase profiling, and Eph receptor activation assays. For further technical details and ordering, please contact Ascent Research.
