The EFNB1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human colorectal adenocarcinoma cell line HT-29. This product provides a heterogeneous pool of cells with targeted disruption of the EFNB1 gene, enabling loss-of-function studies of ephrin-B1 in a genetically unselected background. The polyclonal format avoids clonal bias and is suitable for experiments where population-level effects of gene knockout are of primary interest.
HT-29 is a widely used human colon adenocarcinoma cell line originally isolated from a 44-year-old Caucasian female. These cells serve as a robust model for intestinal epithelial biology and colorectal cancer research. The HT-29 line exhibits an adherent, epithelial-like morphology and is capable of differentiation under appropriate culture conditions, making it a valuable platform for studying tumor cell behavior, drug responses, and signaling pathways relevant to colorectal malignancies.
EFNB1 encodes ephrin-B1, a transmembrane ligand of Eph receptor tyrosine kinases. Upon cell-cell contact, ephrin-B1 engages Eph receptors including EphA1, EphA2, and EphB2, initiating bidirectional signaling. Forward signaling via Eph receptors activates Src family kinases and FAK, directing the small GTPases Rac1 and RhoA to modulate cytoskeletal dynamics and cell adhesion. Reverse signaling through ephrin-B1??s cytoplasmic domain involves PDZ-domain-containing proteins and GIT1, recruiting paxillin and regulating downstream effectors such as ERK1/2, JNK, and Akt. EFNB1 expression is influenced by growth factors including PDGF, TGF-beta, WNT ligands, and FGF. Knockout of EFNB1 disrupts this bidirectional signaling apparatus, offering a loss-of-function model for dissecting ephrin-B1-dependent pathways.
In the HT-29 colorectal cancer model, ephrin-B1-mediated signaling is implicated in tumor cell invasiveness, metastatic potential, and interaction with the tumor microenvironment. Disruption of EFNB1 in these polyclonal cells allows investigation of how loss of ephrin-B1 affects adhesion, migration, proliferation, and signal transduction in a disease-relevant context. The model enables examination of the interplay between Eph-ephrin pathways and other oncogenic pathways such as MAPK/ERK and PI3K-Akt, which are frequently dysregulated in colorectal cancer.
These polyclonal knockout cells are ideally suited for colorectal cancer progression studies, cell migration and invasion assays, and signaling pathway analysis. Researchers can utilize wound healing and transwell migration/invasion assays, western blotting for phosphorylated Eph receptors and ERK, co-immunoprecipitation, immunofluorescence, RT-qPCR, and phospho-ERK analysis. The model also supports drug resistance investigations and neural development models owing to ephrin-B1??s role in axon guidance. For further details, please contact Ascent Research.
