JAG1 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-mediated loss-of-function model in which the JAG1 gene has been disrupted across a polyclonal cell population, eliminating a key ligand of the Notch signaling pathway. This product provides a heterogeneous pool of SK-HEP-1 hepatic adenocarcinoma cells carrying diverse JAG1 knockout alleles, enabling robust interrogation of JAG1-dependent processes without the clonal selection biases inherent to single-cell-derived lines. The polyclonal format preserves the genetic diversity of the edited population, closely mirroring the complexity of tumor cell populations and offering a physiologically relevant tool for studying Notch biology in liver cancer.
The SK-HEP-1 host cell line is a human hepatic adenocarcinoma model derived from ascitic fluid of a patient with liver adenocarcinoma. These cells display an endothelial-like phenotype and have been extensively utilized as a model system for hepatocellular carcinoma, tumor angiogenesis, and epithelial-mesenchymal transition (EMT) studies. Their origin from a metastatic site endows them with aggressive growth characteristics and the capacity to form tubule-like structures in vitro, making them a valuable platform for dissecting the molecular underpinnings of liver cancer progression and vascular mimicry in a controlled experimental setting.
JAG1 encodes a transmembrane ligand that activates Notch receptors (NOTCH1?C4) through direct cell-cell contact, initiating a cascade involving ADAM17-mediated cleavage and subsequent ??-secretase processing (mediated by PSEN1/PSEN2, NCSTN, and APH1) that releases the Notch intracellular domain (NICD). NICD translocates to the nucleus, where it forms a complex with RBPJ/CSL and coactivators such as MAML1 and EP300 to transcriptionally regulate downstream targets including HES1, HES5, HEY1, HEY2, MYC, CCND1, and CDKN1A. JAG1 expression is modulated by upstream stimuli such as TGF-??, VEGF, and HIF1??, and its signaling intersects with the PI3K/Akt/mTOR and JAK/STAT pathways. Disruption of JAG1 in this polyclonal knockout model abolishes ligand-dependent Notch activation, leading to diminished expression of these critical target genes and consequent alterations in cell fate determination, proliferation, and survival.
In the SK-HEP-1 adenocarcinoma context, JAG1-mediated Notch signaling is implicated in maintaining an invasive and pro-angiogenic phenotype. Knockout of JAG1 is expected to attenuate EMT markers such as SNAI1 and TWIST1, reduce survival signals transmitted through BCL2, and impair proliferation driven by MYC and CCND1. This disruption also dampens cross-talk with angiogenic programs by lowering VEGFA induction and alters the cellular response to TGF-??. Consequently, this polyclonal knockout model provides a powerful system for elucidating how JAG1 loss influences hepatic tumor cell plasticity, metastatic potential, and resistance to apoptosis, facilitating the identification of JAG1-dependent vulnerabilities in liver cancer.
This product is ideally suited for a spectrum of research applications, including mechanistic studies of Notch signaling, high-content drug screening for Notch pathway inhibitors, and functional genomics investigations. Typical assays employ Western blotting for JAG1 and NICD, RT-qPCR for HES1/HEY1 transcript levels, Notch reporter assays (CBF1-luciferase), flow cytometric analysis of Notch receptors, and immunofluorescence detection of NICD localization. Co-immunoprecipitation can validate altered JAG1?CNOTCH1 interactions, while proliferation, migration, and apoptosis assays quantify phenotypic consequences. The polyclonal nature makes this tool particularly suitable for CRISPR functional screens and for assessing drug sensitivity to ??-secretase inhibitors in a population setting. For further information or to discuss technical specifications, please contact Ascent Research.
