The IRF3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the IRF3 gene in the HAP1 cell background. This heterogeneous pool offers a loss-of-function model to study interferon regulatory factor 3 (IRF3) without requiring clonal isolation, suitable for population-level analyses of innate immune signaling.
The host HAP1 line is a near-haploid human cell line derived from KBM-7 chronic myelogenous leukemia cells, exhibiting adherent fibroblast-like morphology. Its largely haploid karyotype reduces genetic redundancy, making it an ideal platform for gene disruption studies and enabling clear phenotypic readouts in functional genomics experiments.
IRF3 functions as a central transcription factor in antiviral innate immunity, operating downstream of cytosolic sensors RIG-I, MDA5, and cGAS. Upon activation, the adaptors MAVS and STING relay signals to kinases TBK1 and IKK??, which phosphorylate IRF3. Phosphorylated IRF3 dimerizes and translocates to the nucleus, where it collaborates with NF-??B and coactivators CBP/p300 to drive expression of type I interferons (IFNB1, IFNA) and interferon-stimulated genes such as ISG15, CXCL10, OAS1, and MX1. IRF3 also interacts with IRF7 and ??-catenin, integrating multiple signaling inputs.
In the HAP1 context, IRF3 knockout abrogates type I interferon induction upon viral or nucleic acid challenge, providing a well-defined phenotype. The near-haploid background ensures that a single gene disruption efficiently eliminates function, allowing unambiguous dissection of IRF3-dependent versus IRF7- or NF-??B-mediated pathways and precise assessment of upstream kinase dependencies.
Researchers can apply these polyclonal knockout cells to study innate immune cascades using Western blot for phospho-IRF3, RT-qPCR of IFNB1 and ISGs, IFN?? luciferase reporter assays, and immunofluorescence for IRF3 nuclear localization. The model also supports co-immunoprecipitation to probe IRF3 interactions with TBK1, IKK??, or CBP/p300, and viral challenge assays to explore IRF3-independent antiviral mechanisms. These applications facilitate antiviral drug target validation, autoimmune disease modeling, and investigation of type I interferonopathies. For further details or custom applications, please contact Ascent Research.
