IRF3 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the human colorectal adenocarcinoma HT29 cell line, engineered for disruption of the IRF3 gene. This loss-of-function model eliminates IRF3 expression, enabling investigation of innate immune signaling without clonal biases. The polyclonal format provides a heterogeneous gene-disruption landscape, well-suited for bulk functional genomics and screening applications.
The HT29 cell line originates from a human colorectal adenocarcinoma of female origin and serves as a classic intestinal epithelial model. HT29 cells are employed extensively in cancer biology, drug absorption studies, and mucosal immunity research due to their epithelial characteristics and adaptability to various assay formats, including imaging, reporter gene, and cytokine assays.
IRF3 is a central transcription factor in innate antiviral defense. Pattern recognition receptors RIG-I, cGAS, TLR3, and TLR4 signal via adaptors MAVS, STING, and TRIF to activate kinases TBK1 and IKK??, which phosphorylate IRF3. Phosphorylated IRF3 dimerizes, translocates to the nucleus, and binds ISREs in complex with IRF7 and coactivators CBP/p300. This drives expression of type I interferons (IFN-??, IFN-??), antiviral effectors (ISG15, IFIT1, OAS1), chemokines (CXCL10, RANTES), and pro-apoptotic NOXA. IRF3 also interacts with NF-??B p65, TRAF3, and TRIM21, integrating multiple innate signaling branches.
In HT29 cells, IRF3 knockout abolishes interferon and antiviral gene induction, compromising innate immune responses critical for intestinal epithelial defense. Given HT29??s colorectal cancer origin and the established links between IRF3 signaling and tumor immunity, this model is instrumental for studying how loss of innate sensing impacts tumor biology and the immune microenvironment. IRF3 crosstalk with ??-catenin-driven proliferation further highlights its dual role in immunity and oncogenesis.
These polyclonal knockout cells are validated for key applications: Western blot for IRF3 and phospho-IRF3, RT-qPCR for IFN-?? and ISG15, and luciferase reporter assays for IFN-?? promoter activity. Immunofluorescence can assess IRF3 nuclear translocation, while co-immunoprecipitation examines IRF3-TBK1 interactions. The model supports viral replication studies using VSV-GFP and cytokine profiling by ELISA. It is also appropriate for drug screening targeting the RIG-I/MAVS/IRF3 axis and for exploring colorectal cancer immune evasion mechanisms. For additional product details, please contact Ascent Research.
