The JAG1 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered from the HCT 116 colorectal carcinoma line for loss-of-function studies of JAG1. This heterogeneous pool harbors targeted disruptions in the JAG1 locus, offering a robust model to assess gene-dosage effects without clonal selection artifacts. It is suitable for dissecting JAG1-dependent Notch signaling and its contributions to oncogenic phenotypes.
HCT 116 is a widely characterized colorectal adenocarcinoma cell line with microsatellite instability (MSI-H) and oncogenic mutations in KRAS (G13D) and PIK3CA (H1047R), while retaining TP53 wild-type status. Its epithelial origin and genetic profile make it particularly relevant for studying colorectal cancer progression, therapeutic resistance, and epithelial-mesenchymal transition.
JAG1 functions as a ligand for NOTCH1-4 receptors, initiating cleavage by ADAM10/ADAM17 and ??-secretase to generate the Notch intracellular domain (NICD). NICD complexes with CSL/RBPJ and MAML to transcriptionally activate targets like HES1 and HEY1, which regulate cell proliferation, differentiation, and survival. Upstream regulators include TGF-??, VEGF, HIF1A, TNF-??, and Wnt/??-catenin, while downstream effectors encompass MYC, CCND1, BCL2, SOX9, SNAI1, and VEGFR2. Interactions with MIB1, Fringe proteins, and DLL1/4 further fine-tune Notch signaling dynamics, placing JAG1 at a central node in cell fate determination.
In HCT 116 cells, JAG1-mediated Notch activation sustains tumorigenic traits such as enhanced proliferation, migration, EMT, and angiogenesis. The loss of JAG1 disrupts NICD formation and reduces HES1/HEY1 expression, potentially attenuating these processes. This knockout model leverages the MSI-H and mutant KRAS/PIK3CA context to explore JAG1-dependent crosstalk with Wnt and VEGF pathways, providing insights into colorectal cancer biology and therapeutic vulnerabilities.
Typical applications include western blotting for JAG1 and NOTCH1 cleavage, RT-qPCR for HES1/HEY1, RNA-seq, co-culture assays, migration/invasion studies, proliferation and apoptosis assays, spheroid formation, and drug sensitivity screens. Flow cytometry can monitor stem cell marker changes. These cells enable comprehensive investigation of Notch signaling in colorectal cancer, tumor microenvironment interactions, and JAG1-targeted drug discovery. For further details, contact Ascent Research.
