The IL1R1 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human colorectal adenocarcinoma cell line HT29, in which the interleukin-1 receptor type 1 (IL1R1) gene has been disrupted. This polyclonal pool provides a heterogeneous loss-of-function model enabling robust assessment of IL-1 signaling in an epithelial colorectal cancer background. Generated via CRISPR/Cas9-mediated gene disruption, the cells serve as a powerful tool for dissecting IL1R1-dependent mechanisms without the constraints of clonal selection.
The HT29 host cell line originates from a 44-year-old female with colorectal adenocarcinoma and exhibits adherent epithelial morphology. Widely employed in cancer biology studies, HT29 cells retain key features of intestinal epithelial cells and are responsive to inflammatory cytokines, making them an ideal platform for investigating the interplay between inflammation and tumorigenesis in the colorectal epithelium.
IL1R1 encodes the primary receptor for the pro-inflammatory cytokines interleukin-1 alpha (IL-1??) and interleukin-1 beta (IL-1??). Ligand binding triggers receptor complex formation with the IL-1 receptor accessory protein (IL-1RAcP) and recruitment of the adaptor MyD88, leading to activation of IRAK4 and IRAK1. These kinases associate with TRAF6, which in turn activates downstream pathways including NF-??B and MAPK cascades (ERK, p38, JNK). Consequently, IL1R1 signaling transcriptionally upregulates a battery of inflammatory mediators such as IL-6, TNF-??, and COX-2, as well as the transcription factor AP-1. The pathway is tightly controlled by the endogenous antagonist IL-1Ra. In the knockout cells, IL-1 stimulation fails to induce NF-??B nuclear translocation or phosphorylation of p65 and MAPKs, thereby abrogating the expression of key pro-inflammatory target genes.
In the context of HT29 colorectal adenocarcinoma cells, IL1R1-mediated signaling contributes to inflammation-driven processes influencing tumor cell proliferation, migration, and survival. The loss of IL1R1 in this background allows researchers to uncouple the autocrine and paracrine effects of IL-1 from other oncogenic pathways active in colorectal cancer. This knockout model is particularly relevant for studying how tumor-derived epithelial cells respond to inflammatory cues within the tumor microenvironment and for evaluating the impact of IL-1 signaling blockade on malignant phenotypes.
The IL1R1 Knockout HT29 Polyclonal Cells are well-suited for a variety of assays including Western blotting for NF-??B, phospho-p65, and phospho-MAPKs; RT-qPCR analysis of pro-inflammatory transcripts such as IL-6, IL-8, and COX-2; NF-??B luciferase reporter assays; and ELISA quantification of secreted cytokines. Functional studies can encompass cell proliferation, migration/invasion, and apoptosis assays, alongside immunofluorescence detection of NF-??B subcellular localization. These applications support investigations into IL-1 signaling in colorectal cancer, inflammation-driven tumorigenesis, anti-inflammatory drug screening, and cytokine signaling analysis. For further information, please contact Ascent Research.
