The AZI2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of HT29 cells harboring targeted disruption of the AZI2 gene. This loss-of-function model eliminates functional AZI2 protein, enabling investigation of its roles in innate immune signaling, autophagy, and inflammation. As a polyclonal pool, the cells provide a robust system free from clonal artifacts, suitable for studying pathway-level effects.
The HT29 cell line is a human colorectal adenocarcinoma of epithelial origin, widely used for intestinal epithelial studies and cancer research. Its epithelial phenotype, including polarized monolayer formation and mucin expression, provides a relevant physiological context for examining mucosal innate immunity and tumor microenvironment interactions.
AZI2 acts as an adaptor scaffolding TBK1 and IKK?? kinases downstream of pattern recognition receptors such as RIG-I, MDA5, and cGAS. Upon stimulation by viral dsRNA, cGAMP, LPS, or poly(I:C), AZI2 facilitates phosphorylation of IRF3/IRF7 and NF-??B p65, driving type I interferon and proinflammatory cytokine expression. It also regulates autophagy, reflected by LC3 lipidation and p62 degradation. Key interacting partners include TBK1, IKK??, STING, TRAF3, and MAVS.
Within the HT29 colorectal cancer context, AZI2 knockout permits dissection of innate immune pathway contributions to tumor cell biology. Colorectal malignancies frequently display dysregulated NF-??B and interferon responses that affect growth and immune evasion. This model thus enables precise analysis of AZI2-dependent IRF3/NF-??B activation, cytokine secretion, and autophagy in a cancer-relevant epithelial background.
Applications include Western blotting for phospho-IRF3 and phospho-p65, RT-qPCR for IFN-??/ISGs, luciferase reporter assays, co-immunoprecipitation of AZI2 complexes, and immunofluorescence for IRF3 nuclear translocation. Functional assays cover cell viability, migration, apoptosis, cytokine ELISA, viral replication, and autophagy flux (LC3-II with chloroquine). This versatile model supports antiviral signaling research, cancer immunology, and drug target validation. For technical inquiries, contact Ascent Research.
