The EFNB1 Knockout SK-HEP-1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt expression of the EFNB1 gene in the human SK-HEP-1 cell line. This pool of edited cells provides a heterogeneous loss-of-function model for studying ephrin-B1-dependent signaling without clonal selection, making it suitable for applications where polyclonal knockout representation is preferred. The cell population is generated through CRISPR/Cas9-mediated gene disruption, resulting in a mixed population with targeted mutations in EFNB1.
The host cell line, SK-HEP-1, is a human hepatic adenocarcinoma-derived cell line with an epithelial morphology. Originally isolated from a liver adenocarcinoma patient, SK-HEP-1 cells are extensively used in hepatocellular carcinoma research, drug metabolism studies, and cancer biology investigations. This adherent cell line offers a robust in vitro platform for examining liver cancer progression, metastatic behavior, and therapeutic responses in a genetically defined background.
EFNB1 encodes ephrin-B1, a transmembrane ligand for Eph receptor tyrosine kinases, which mediates bidirectional signaling into both the ligand-expressing cell (reverse signaling) and the receptor-expressing cell (forward signaling). Ephrin-B1 reverse signaling is activated by binding to Eph receptors such as EPHB2 and involves downstream effectors including GRB4, PDZ-RGS3, and the Rho GTPases RHOA and RAC1, ultimately modulating actin cytoskeleton dynamics through ROCK, LIMK, and Cofilin. Additionally, ephrin-B1 engagement can activate the MAPK/ERK pathway (ERK1/2) and focal adhesion kinase (FAK) signaling, while its expression is transcriptionally regulated by upstream factors such as HOXA5, RUNX2, TGFB1, and WNT signaling. Ephrin-B1 also interacts with ADAM10 protease and clathrin-mediated endocytosis machinery, linking it to receptor shedding and signal attenuation.
In the SK-HEP-1 liver cancer model, knockout of EFNB1 disrupts bidirectional Eph-ephrin signaling, impairing cell adhesion and migration, two processes critical for tumor invasion and metastasis. Loss of ephrin-B1 reduces Rho GTPase-mediated cytoskeletal dynamics and downregulates oncogenic MAPK/ERK pathway activity, which may attenuate the malignant phenotype of these hepatocellular carcinoma cells. This polyclonal knockout pool allows researchers to assess the overall impact of ephrin-B1 deficiency on liver cancer cell behavior without clonal artifacts, making it a valuable tool for mechanistic studies in hepatic oncology.
This product is well-suited for a range of experimental applications, including tumor invasion and metastasis assays, liver cancer drug screening, and cell-cell communication studies. Researchers can employ techniques such as Western blotting, RT-qPCR, immunofluorescence, migration and invasion assays, phospho-signaling analysis, and co-immunoprecipitation to characterize the molecular consequences of EFNB1 disruption. The polyclonal knockout population is particularly advantageous for pooled functional screens and bulk signaling studies. For additional information or technical support, please contact Ascent Research.
