The EFNB1 Knockout HEK293T Polyclonal Cells product offers a CRISPR/Cas9-edited polyclonal knockout cell population targeting the EFNB1 gene in the HEK293T human embryonic kidney cell background. This loss-of-function model is generated through CRISPR/Cas9-mediated gene disruption, resulting in a heterogeneous pool of cells with targeted gene knockout. Suitable for studying ephrin-B1 function, this polyclonal population provides a reproducible tool for investigating EFNB1-dependent signaling mechanisms without the need for single-cell clonal isolation.
HEK293T cells are a widely utilized human embryonic kidney epithelial cell line derived from the HEK293 parental line and stably expressing the SV40 large T antigen. This characteristic enables episomal replication of plasmids containing the SV40 origin, contributing to high transfection efficiency and robust protein expression. HEK293T cells are a preferred host for viral packaging, protein production, and a broad range of cell biology assays, making them an ideal platform for interrogating gene function via CRISPR-based knockout strategies.
EFNB1 encodes ephrin-B1, a transmembrane ligand for EphB receptor tyrosine kinases, which mediates bidirectional signaling critical for cell adhesion, repulsion, and migration during development. Forward signaling through EphB receptors activates downstream effectors such as Src family kinases, Rac1, and Cdc42, while reverse signaling via the ephrin-B1 intracellular domain recruits PDZ domain proteins like syntenin and PICK1, and adaptors including Grb4 (NCK2), linking to Rho family GTPase regulation and MAPK/ERK pathway activation. Ephrin-B1 is transcriptionally regulated by Sp1 and NF-Y, and its signaling is modulated by EphB1, EphB2, and EphB3 receptor interactions. Disruption of EFNB1 abolishes both forward and reverse signaling, leading to altered Rho GTPase activity and impaired cytoskeletal dynamics.
In the HEK293T background, EFNB1 knockout creates a null background for ephrin-B1, enabling dissection of Eph/ephrin signaling pathways without endogenous ligand interference. This model recapitulates molecular hallmarks of craniofrontonasal syndrome (CFNS), an X-linked disorder caused by EFNB1 mutations, characterized by abnormal skeletal development and cell migration defects. The absence of ephrin-B1 in these polyclonal cells provides a valuable system for exploring the role of EFNB1 in cancer metastasis, where ephrin-B1 influences tumor cell invasion and adhesion dynamics through modulation of Rac1, Cdc42, and Src signaling.
These polyclonal knockout cells are suitable for a wide array of experimental applications, including CFNS pathology studies, investigation of Eph/ephrin reverse signaling, and cancer cell migration and invasion assays. Representative techniques include western blotting to confirm loss of ephrin-B1 protein, RT-qPCR for EFNB1 mRNA quantification, immunofluorescence for subcellular localization, scratch wound-healing assays, transwell invasion assays, co-immunoprecipitation with EphB receptors, and phospho-signaling arrays. The polyclonal nature allows for the study of bulk population effects, facilitating drug screening for Ephrin-B1 pathway modulators and developmental biology research. For further information, please contact Ascent Research.
