The IRAK1 Knockout HCT 116 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the human IRAK1 gene. This mixed population arises from targeted disruption of IRAK1 in the HCT 116 colorectal carcinoma cell line, providing a physiologically relevant model to interrogate interleukin-1 receptor (IL-1R) and Toll-like receptor (TLR) signaling. The polyclonal format avoids the phenotypic artifacts of clonal selection while offering a robust background for pathway analysis, drug screening, and functional genomics.
The host HCT 116 cell line is an extensively characterized epithelial model of colorectal cancer, harboring a KRAS G13D driver mutation and wild-type TP53. Notably, these cells exhibit microsatellite instability (MSI), a hallmark of defective DNA mismatch repair commonly observed in colorectal tumors. The genetic context of activated KRAS and intact p53 enables investigation of oncogenic signaling interactions with innate immune pathways, making HCT 116 a relevant platform for studying inflammation-driven cancer progression.
IRAK1 encodes a serine/threonine kinase that functions downstream of MyD88-dependent IL-1R and TLR signaling. Upon ligand stimulation, IRAK1 is recruited to the MyD88 adaptor complex and phosphorylated by IRAK4, facilitating its association with TRAF6. This interaction promotes TAK1-dependent activation of the IKK complex and MAP kinases, culminating in NF-??B and AP-1-mediated transcription of pro-inflammatory cytokines. IRAK1 also interacts with regulatory proteins such as Pellino-1 and Tollip, which modulate signal amplitude and duration. Thus, IRAK1 represents a central node in innate immune signaling, linking receptor activation to transcriptional responses.
In the HCT 116 colorectal cancer background, IRAK1 disruption provides a unique tool to dissect the role of TLR/IL-1R signaling in tumor biology. Given the cell line??s MSI status and KRAS mutation, this model enables exploration of how inflammatory signals intersect with oncogenic pathways to influence cell proliferation, survival, and chemoresistance. Moreover, it allows assessment of IRAK1 dependency in a context where NF-??B and MAPK pathways are frequently hyperactive, aiding in validation of therapeutic targets for cancers with an inflammatory microenvironment.
Researchers can employ this knockout population in a variety of advanced assays, including CRISPR/Cas9-mediated gene disruption analysis by Western blot for IRAK1 protein levels, RT-qPCR for mRNA quantification, and NF-??B luciferase reporter assays to measure pathway activity. ELISA-based detection of secreted cytokines such as IL-6 and IL-8, phospho-signaling analysis of downstream effectors like p-IRAK1, p-p65, and p-JNK, and cell viability or drug sensitivity studies with IRAK1 inhibitors further expand its utility. For additional technical specifications or support, please contact Ascent Research.
