The IL1RN Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Jurkat T-lymphocyte line, engineered to disrupt the endogenous IL1RN gene encoding interleukin-1 receptor antagonist (IL-1Ra). This polyclonal pool carries heterogeneous loss-of-function mutations introduced at the target locus, generating a reliable loss-of-function model for investigating IL-1 signaling without the influence of clonal selection. The cells are suitable for studying the functional consequences of IL-1Ra ablation in a T-cell context.
Jurkat cells originate from an acute T-cell leukemia and serve as an immortalized CD4+ T-cell model, widely employed to probe T-cell receptor (TCR) signaling, IL-2 production, and activation-induced gene regulation. They grow in suspension and maintain expression of the TCR/CD3 complex. Although basal IL-1R1 levels are low, inflammatory cues such as TNF or phorbol esters can upregulate pathway components, making them a tractable host for examining IL-1-dependent responses in lymphocytes.
The IL1RN product, IL-1Ra, is a competitive antagonist that binds IL1R1 without recruiting IL1RAP, thereby blocking IL-1??/??-driven signaling. In wild-type cells, this prevents MyD88?CIRAK1?CTRAF6 assembly and downstream NF-??B (via IKBKB and NFKB1) and MAPK (MAPK3/MAPK1) pathways, curbing transcription of IL6, TNF, IL8, and PTGS2. IL1RN itself is induced by NF-??B and AP-1 upon exposure to LPS, TNF, or IL-1, forming a negative feedback loop. Knockout of IL1RN removes this brake, allowing unopposed IL-1 receptor activation and amplified inflammatory gene expression.
In the Jurkat background, this knockout unveils the role of IL-1Ra in modulating T-cell inflammatory responses. Without IL-1Ra, exogenous IL-1?? hyperactivates NF-??B and MAPK cascades, which can synergize with TCR signals to reshape cytokine profiles and cell fate decisions. The model is valuable for exploring how chronic IL-1 exposure influences leukemic T-cell biology and for dissecting IL-1?CTCR crosstalk in the absence of feedback inhibition.
Applications include ELISA measurement of IL-6/TNF secretion after IL-1?? stimulation, Western blotting for phospho-p65 and phospho-p38, and RT-qPCR analysis of inflammatory gene induction. The cells support NF-??B luciferase reporter assays, flow cytometric detection of IL-1R1, and co-immunoprecipitation studies. They are also suitable for screening small-molecule inhibitors of the IL-1 pathway, such as IRAK1 or TAK1 blockers, in the context of cancer-associated inflammation. For further information or custom projects, contact Ascent Research.
