IL1R1 Knockout 786-O Polyclonal Cells represent a CRISPR/Cas9-engineered polyclonal population derived from the human renal cell carcinoma line 786-O, in which the IL1R1 gene has been disrupted to eliminate functional expression of the type I interleukin-1 receptor. This polyclonal knockout product provides a heterogeneous mixture of edited cells, offering a robust loss-of-function model that avoids artifacts associated with single-cell cloning while enabling pooled screening and population-level analyses of IL-1 signaling.
The 786-O host cell line is a well-characterized model of human clear cell renal cell carcinoma (ccRCC), harboring a biallelic loss-of-function mutation in the von Hippel?CLindau (VHL) tumor suppressor gene. This defect leads to constitutive stabilization of hypoxia-inducible factors (HIFs), driving pseudohypoxic gene expression programs that contribute to tumorigenesis, metabolic reprogramming, and angiogenesis. 786-O cells are extensively employed in studies of ccRCC biology, including the interplay between oncogenic signaling and the inflammatory tumor microenvironment.
The IL1R1 gene encodes the type I interleukin-1 receptor, which upon binding the agonists IL1A or IL1B, associates with the co-receptor IL1RAP and the adaptor MYD88. This triggers signaling via the kinases IRAK4 and IRAK1, leading to activation of TRAF6 and TAK1. Activated TAK1 directs phosphorylation of IKBKB, resulting in NF-??B translocation via degradation of the inhibitor NFKBIA, and also activates MAPK pathways including JNK and p38. These pathways drive expression of pro-inflammatory and angiogenic effectors including IL6, CXCL8, and VEGFA. The pathway is negatively regulated by the decoy receptor IL1R2, the antagonist IL1RN, and adaptors like TOLLIP and SIGIRR.
In the context of 786-O cells, loss of IL1R1 function provides a precise tool to dissect the contribution of IL-1-mediated signaling to the pro-tumorigenic inflammatory network of ccRCC. With constitutive HIF activation due to VHL deficiency, these cells exhibit elevated baseline expression of angiogenic factors that can be further potentiated by IL-1. Disruption of IL1R1 abrogates IL-1-dependent augmentation of tumor-promoting molecules such as VEGFA, IL-6, IL-8, and MMP9, enabling researchers to decouple extrinsic cytokine stimulation from intrinsic oncogenic signaling. This model is valuable for investigating IL-1-driven NF-??B and MAPK crosstalk with HIF pathways and for assessing chemokine-mediated immune cell recruitment and matrix remodeling in the kidney cancer microenvironment.
This polyclonal IL1R1 knockout cell product is designed for detailed functional and pharmacological studies. Western blotting monitors changes in phospho-p65 (RELA), phospho-p38, and phospho-JNK as pathway readouts. RT-qPCR and ELISA enable profiling of IL-1-dependent transcriptional responses, including IL6, IL8, and CXCL8. NF-??B activity is quantified via luciferase reporter assays, and flow cytometry validates loss of IL1R1 surface expression. Co-immunoprecipitation studies impaired IL1R1?CIL1RAP complex formation, while migration or invasion assays under IL-1?? stimulation assess cytokine-driven metastatic behavior. These cells are also suitable for inhibitor screening and RNA-seq transcriptomic profiling. For further technical inquiries, please contact Ascent Research.
