The IL27 Knockout NCI-H1299 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population derived from the NCI-H1299 human lung carcinoma epithelial cell line, engineered for targeted disruption of the IL27 gene. This product provides a heterogeneous pool of knockout cells exhibiting loss-of-function for the p28 subunit of IL-27, enabling researchers to interrogate autocrine and paracrine cytokine signaling networks in a non-small cell lung cancer (NSCLC) context without clonal selection artifacts. The polyclonal format preserves the inherent genetic diversity of the parental line while abolishing IL27 expression, making it suitable for pooled functional screens and robust population-level analyses of tumor-immune interactions.
The NCI-H1299 host cell line was originally derived from a metastatic lymph node of a 43-year-old male patient with lung carcinoma and has since become a widely employed model for NSCLC research. As an epithelial cell line lacking normal p53 function, NCI-H1299 cells exhibit aggressive growth characteristics and are extensively used to study tumor cell biology, drug resistance mechanisms, and oncogenic signaling pathways. This well-characterized background allows focused examination of IL27-mediated effects on proliferation, migration, and immune modulatory phenotypes relevant to lung cancer progression.
IL-27 is a heterodimeric cytokine composed of the p28 subunit (encoded by IL27) and EBI3 (IL-27B), which signals through a receptor complex consisting of IL27RA (WSX-1) and gp130 (IL6ST). Upon ligand engagement, the receptor-associated kinases JAK1 and JAK2 are activated, leading to phosphorylation of STAT1 and STAT3, which are the principal downstream effectors. This signaling cascade transcriptionally induces T-bet (TBX21) to drive Th1 differentiation and also upregulates the anti-inflammatory cytokine IL-10 and the feedback inhibitor SOCS1, while simultaneously repressing Th17 responses. Upstream activators such as TLR4 agonists (LPS), IFN-??, and CD40 engagement trigger IL27 expression, linking innate immune stimuli to adaptive immune modulation. Additionally, IL-27 has been implicated in regulating CD274 (PD-L1) expression, connecting it to immune checkpoint pathways.
In the lung cancer microenvironment, IL-27 exerts pleiotropic effects that can either promote or restrain tumor immunity depending on the cellular context. NSCLC cells may exploit IL-27 signaling to induce PD-L1 expression, thereby facilitating immune evasion, or to modulate the balance between effector T-cell subsets and regulatory IL-10-producing cells. The NCI-H1299 knockout model allows dissection of these tumor-intrinsic functions of IL-27, separating the cytokine??s effects on malignant epithelial cells from its roles in infiltrating immune cells. Researchers can thus investigate how loss of IL27 alters downstream STAT1/STAT3 phosphorylation, SOCS1 induction, and transcriptional programs controlling immune-related gene expression within the tumor cells themselves.
This knockout tool is ideal for a broad spectrum of experimental applications, including co-culture systems with primary T cells to evaluate altered tumor?Cimmune crosstalk, cytokine profiling via ELISA for IL-27 and multiplexed secretome analysis, Western blotting for phospho-STAT1/STAT3, and RT-qPCR quantification of downstream targets such as IL10, SOCS1, and TBX21. Flow cytometric assessment of PD-L1 surface expression further enables correlation with immune checkpoint modulation. Proliferation and migration assays under defined cytokine conditions can reveal functional consequences of IL27 loss. These polyclonal knockout cells thus serve as a versatile platform for target validation studies, drug response screening against IL-27 pathway inhibitors, and mechanistic interrogation of JAK-STAT-mediated tumor-autonomous immune regulation. For additional information or technical support, please contact Ascent Research.
