The JAG2 Knockout KYSE-30 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human esophageal squamous cell carcinoma (ESCC) cell line KYSE-30. This product provides a heterogeneous pool of cells with target-gene disruption at the JAG2 locus, enabling loss-of-function studies in a physiologically relevant ESCC background. As a polyclonal population, these cells retain the genetic diversity of the original knockout pool, making them suitable for functional screens, pooled analyses, and studies where clonal artifacts may be a concern. The knockout was generated using CRISPR/Cas9 technology to disrupt JAG2, eliminating expression of the full-length protein and abrogating its role as a Notch ligand. These cells are ideal for investigating JAG2-dependent signaling in cancer biology, drug resistance, and cell fate regulation.
KYSE-30 is a well-characterized human ESCC cell line originally established from a moderately differentiated esophageal squamous cell carcinoma. It is widely used as a model for ESCC, a prevalent and aggressive malignancy with poor prognosis. KYSE-30 cells exhibit epithelial morphology, express characteristic squamous cell markers, and harbor genetic alterations common in ESCC, including TP53 mutations. They are highly tumorigenic in xenograft models and display features of the epithelial-mesenchymal transition (EMT) upon stimulation. This cell line serves as a robust platform for studying oncogenic signaling, metastatic mechanisms, and therapeutic responses relevant to esophageal cancer.
JAG2 encodes Jagged-2, a canonical Notch ligand that activates NOTCH1 and NOTCH3 receptors on adjacent cells. Upon ligand-receptor interaction, sequential proteolytic cleavages mediated by ADAM17 and the ??-secretase complex (including PSEN1) release the Notch intracellular domain (NICD). NICD translocates to the nucleus, where it forms a transcriptional activation complex with the DNA-binding protein RBPJ and co-activator MAML1. This complex drives expression of downstream targets such as HES1, HEY1, HEY2, MYC, and CCND1, thereby controlling cell proliferation, differentiation, and survival. JAG2 expression is regulated by upstream factors including EGF, TGF-??, HIF1A, and the related ligand JAG1, positioning it at a key node in the Notch signaling network. In cancer contexts, JAG2-mediated Notch activation can promote EMT, stemness, and metastatic dissemination.
In KYSE-30 cells, JAG2 is frequently overexpressed and contributes to the malignant phenotype by sustaining Notch pathway activation. Disruption of JAG2 in this model allows researchers to dissect the ligand-specific contributions of Jagged-2 versus other Notch ligands (e.g., JAG1, DLL4) to ESCC progression. The polyclonal knockout pool enables assessment of heterogeneous responses, better mimicking the tumor microenvironment. These cells can be used to evaluate changes in cell growth, migration, invasion, and drug sensitivity attributable to JAG2 loss. Moreover, they provide a syngeneic background for co-culture experiments with NOTCH1-expressing reporter cells to measure Notch activity directly. By comparing wild-type and JAG2-disrupted populations, researchers can delineate downstream signaling alterations and identify novel therapeutic targets in ESCC.
The JAG2 Knockout KYSE-30 Polyclonal Cells support a broad range of research applications. They are particularly suited for Notch signaling dissection using RT-qPCR to quantify HES1 and HEY1 expression, western blotting for cleaved NOTCH1, and flow cytometry to confirm loss of surface JAG2. Functional assays such as proliferation (MTS/CCK-8), migration, and invasion can be employed to assess JAG2-dependent aggressiveness. Transcriptomic profiling via RNA-seq can reveal global gene expression changes, while co-culture Notch reporter assays provide direct readouts of signaling output. These cells also enable studies of drug resistance mechanisms, cancer stem cell biology, and EMT regulation in ESCC. For additional information or customized applications, please contact Ascent Research.
