The IRAK1 Knockout HT29 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal population derived from the human colorectal adenocarcinoma cell line HT29, featuring targeted disruption of the IRAK1 gene. This loss-of-function model enables dissection of IRAK1-dependent signaling in a well-characterized intestinal epithelial context. The polyclonal knockout pool retains genetic heterogeneity while abrogating IRAK1 expression, providing a robust system for genotype-phenotype analyses without single-cell cloning.
HT29 cells were established from a primary colon adenocarcinoma of a 44-year-old Caucasian female and display an undifferentiated phenotype under standard conditions, with the capacity for enterocytic differentiation upon induction. These cells are widely used as an intestinal epithelial model and express functional Toll-like receptors (TLRs) and interleukin-1 receptors (IL-1Rs), making them highly suitable for studying innate immune signaling pathways.
IRAK1 is a serine/threonine kinase that acts as a key adaptor downstream of MyD88 in TLR/IL-1R signaling. Upon ligand binding (e.g., IL-1??, LPS, CpG DNA), IRAK1 is recruited and phosphorylated by IRAK4, then associates with TRAF6 and Pellino proteins. This leads to TRAF6-driven TAK1 activation, which phosphorylates the IKK complex, resulting in NF-??B translocation and transcriptional activation of pro-inflammatory cytokines such as TNF-??, IL-6, and IL-8. In parallel, TAK1 activates MAPK cascades leading to AP-1 induction. IRAK1 activity is modulated by interacting factors including Tollip and TAB2, and it functions as a central mediator coupling receptor activation to inflammatory gene expression.
In the HT29 colorectal adenocarcinoma background, IRAK1 knockout allows investigation of the interplay between innate immune signaling and cancer cell behavior. HT29 cells respond to inflammatory stimuli, so loss of IRAK1 can elucidate its role in tumor-associated inflammation, cytokine production, and treatment resistance. This model also permits analysis of IRAK1-dependent NF-??B effects on proliferation and differentiation, with relevance to colitis-associated cancer and inflammatory bowel disease.
Representative applications include western blotting for IRAK1 and phospho-IRAK1, RT-qPCR for cytokine transcripts, NF-??B reporter assays, ELISA for secreted TNF-?? and IL-6, and flow cytometry for surface receptors. Co-immunoprecipitation can assess disrupted MyD88-IRAK1 interactions, and viability assays can evaluate drug sensitivity. This product is ideal for studying innate immunity, colorectal cancer, inflammation, and NF-??B signaling. For further technical details, please contact Ascent Research.
