This product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the KYSE-150 human esophageal squamous cell carcinoma line, engineered for disruption of the CD274 gene. The knockout model eliminates expression of programmed death-ligand 1 (PD-L1), an immune checkpoint molecule. The polyclonal nature preserves heterogeneous genetic backgrounds while targeting the locus of interest, providing a physiologically relevant loss-of-function system. This pool is suitable for studying PD-L1-dependent immune modulation without clonal artifacts.
KYSE-150 cells were originally established from a poorly differentiated esophageal squamous cell carcinoma and serve as a widely used in vitro model for esophageal cancer research. These adherent epithelial cells retain key characteristics of the tumor subtype, including relevant oncogenic signaling networks. The KYSE-150 line is responsive to inflammatory stimuli such as interferon-gamma (IFN-??), which upregulates CD274 expression via JAK/STAT signaling, making it an appropriate host for investigating PD-L1 biology.
CD274 encodes PD-L1, a transmembrane glycoprotein that functions as a critical immune checkpoint ligand. PD-L1 interacts with the PD-1 receptor on T cells and with CD80, delivering inhibitory signals that suppress T-cell activation and effector functions. PD-1 engagement recruits the phosphatase SHP-2, which dephosphorylates proximal T-cell receptor (TCR) signaling molecules, including ZAP70, thereby attenuating downstream PI3K/AKT and MAPK cascades. Expression of CD274 is regulated by multiple factors, including IFN-?? through JAK/STAT, as well as MYC, ALK, and TNF-??, linking oncogenic and inflammatory pathways to immune evasion.
In KYSE-150 esophageal carcinoma cells, endogenous PD-L1 contributes to immune escape by inhibiting T-cell activity within the tumor microenvironment. Disruption of CD274 in this model system ablates PD-L1 expression, potentially restoring T-cell-mediated antitumor responses. This knockout model thus enables dissection of PD-L1-dependent immune suppression mechanisms in the context of esophageal malignancies. Furthermore, it serves as a tool to evaluate the functional consequences of blocking the PD-1/PD-L1 axis, complementing studies with therapeutic antibodies.
These polyclonal knockout cells are intended for a broad range of immuno-oncology applications. Researchers can employ them in co-culture assays with human T cells to assess T-cell proliferation, cytotoxicity, and cytokine release by ELISA or flow cytometry. The knockout model is also valuable for validating antibody-based PD-L1 detection reagents and screening small-molecule inhibitors targeting the PD-L1 axis. Western blotting, RT-qPCR, and immunofluorescence can confirm the loss of PD-L1 protein and mRNA. For further information or technical support, please contact Ascent Research.
