The IL1RAP Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, engineered to disrupt the IL1RAP gene. This heterogeneous cell pool lacks expression of the interleukin-1 receptor accessory protein (IL-1RAcP), providing a reliable loss-of-function model for investigating IL1RAP-dependent signaling in an epithelial colorectal cancer background.
The HT29 line was established from a 44-year-old female with Dukes?? stage C colorectal adenocarcinoma and displays epithelial morphology. It carries well-characterized oncogenic mutations in APC, BRAF (V600E), and TP53, while retaining wild-type KRAS, and can differentiate under metabolic stress, establishing it as a robust model for colorectal tumor biology.
IL1RAP encodes IL-1RAcP, a co-receptor required for pro-inflammatory signaling by IL-1??, IL-1??, and IL-33. IL-1RAcP heterodimerizes with ligand-binding receptors IL-1R1 and ST2, recruits MyD88, IRAK1, IRAK4, and TRAF6, and triggers activation of the NF-??B and MAPK (ERK, JNK) pathways. This results in transcriptional upregulation of cytokines such as IL-6, IL-8, and TNF, as well as COX-2 and matrix metalloproteinases. Negative modulators include TOLLIP and SIGIRR.
Disruption of IL1RAP in HT29 cells prevents assembly of functional IL-1R1/IL-1RAcP and ST2/IL-1RAcP co-receptor complexes, abrogating downstream NF-??B and AP-1 activation and pro-inflammatory cytokine production. Given the prominence of IL-1 and IL-33 within the colorectal tumor microenvironment, these cells offer a clinically relevant system to examine IL1RAP??s impact on tumor cell proliferation, survival, and inflammatory crosstalk.
The IL1RAP Knockout HT29 Polyclonal Cells support a broad range of research applications, including dissecting IL-1 and IL-33 signaling mechanisms in colon cancer, validating IL1RAP as a therapeutic target, and screening pathway inhibitors. Typical experimental workflows include western blotting for phosphorylated NF???B and MAPK, RT?qPCR for IL?6 and IL?8 transcripts, ELISA for secreted cytokines, and NF???B luciferase reporter assays. Flow cytometry can assess surface IL?1R1 expression, while functional assays evaluate proliferation and apoptosis under IL?1 stimulation. RNA?seq transcriptomics and drug sensitivity profiling further enable comprehensive pathway analysis. For additional information or to discuss specific experimental needs, please contact Ascent Research.
