The EFNB1 Knockout HeLa Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of HeLa cells carrying a targeted disruption of the EFNB1 gene. This loss-of-function model enables investigation of ephrin-B1-dependent signaling, adhesion, and migration in a widely used human cervical adenocarcinoma background. The polyclonal format provides a diverse knockout pool suitable for pooled functional studies without requiring single-cell cloning.
HeLa cells are a well-characterized human cervical adenocarcinoma epithelial line immortalized through HPV-18 integration. They serve as a robust model for studying cancer cell biology, including proliferation, invasion, and cytoskeletal regulation. Their epithelial origin and expression of relevant Eph receptors make them particularly suitable for examining the role of ephrin-B1 in tumor cell behavior and Eph/ephrin signaling.
EFNB1 encodes ephrin-B1, a transmembrane ligand for Eph receptor tyrosine kinases such as EphB2, EphB3, and EphB4. Ephrin-B1 mediates bidirectional signaling: forward signaling through EphB receptors activates Src family kinases, focal adhesion kinase (FAK), and Rho GTPases (RhoA, Rac1, Cdc42), while reverse signaling into the ligand-expressing cell involves adaptor proteins like Grb4 and PDZ domain-containing proteins including GRIP1 and syntenin. These pathways converge on MAPK/ERK cascades and cytoskeletal reorganization. Upstream regulators of EFNB1 expression include MSX2 transcription factor, HOX genes, and Wnt/??-catenin signaling.
In HeLa cells, knockout of EFNB1 disrupts Eph-ephrin bidirectional signaling, impairing cell adhesion, migration, and actin cytoskeletal dynamics. This can affect integrin-mediated adhesion and Rho GTPase-driven protrusion, potentially reducing the invasive capacity of the cancer cells. The model allows dissection of ephrin-B1 reverse signaling independent of Eph receptor forward signaling, providing a valuable tool for studying cancer progression and metastatic dissemination.
Typical applications include cancer cell migration and invasion assays using Boyden chambers, cell adhesion assays, immunofluorescence analysis of actin cytoskeleton organization, and phospho-signaling profiling of Src, FAK, and ERK1/2. Western blotting and RT-qPCR can confirm EFNB1 disruption, while flow cytometry enables assessment of Eph receptor surface expression. This polyclonal knockout pool is also suited for drug screening aimed at Eph/ephrin-targeted therapies. For further product information, please contact Ascent Research.
