The IFI16 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from HEK293T human embryonic kidney cells. This loss-of-function model features targeted disruption of the IFI16 gene across a diverse cell pool, eliminating IFI16 protein expression. The polyclonal format captures genetic heterogeneity, enabling population-level functional studies without clonal bias, and provides a stable alternative to transient knockdown approaches.
HEK293T cells, a derivative of HEK293 transformed with adenovirus 5 DNA, constitutively express the SV40 large T-antigen, which enhances plasmid replication and protein production. They are widely employed for ectopic gene expression, lentivirus production, and biochemical assays, making them a robust and experimentally tractable host for signaling research.
IFI16 functions as a cytosolic DNA sensor that activates dual innate immune pathways. Upon double-stranded DNA recognition, IFI16 recruits STING and triggers TBK1-dependent IRF3 phosphorylation, inducing type I interferon transcription, including IFNB1. Simultaneously, IFI16 assembles the AIM2 inflammasome with ASC and caspase-1, leading to IL-1? and IL-18 maturation and pyroptosis. IFI16 also interacts with p53 and BRCA1, connecting innate immunity to apoptosis and cell cycle control. Its expression is upregulated by interferon-gamma and type I interferons via STAT1 and IRF1, reinforcing antiviral responses.
In the HEK293T background, IFI16 knockout allows dissection of DNA-sensing mechanisms without endogenous IFI16 interference. These cells support reconstitution of STING signaling through transfection and contain the downstream machinery for NF-?B and apoptosis. The SV40 large T-antigen neutralizes p53, simplifying study of p53-independent IFI16 functions. This model is ideal for investigating STING-TBK1-IRF3 signaling and AIM2 inflammasome activation in a clean genetic system.
Applications include analysis of interferon induction, inflammasome assembly, and antiviral responses. Researchers can perform western blotting for phospho-TBK1 and phospho-IRF3, RT-qPCR for IFNB1, caspase-1 activity assays, and ELISA for IL-1?. Co-immunoprecipitation of STING or ASC, immunofluorescence microscopy, and Annexin V flow cytometry are amenable. RNA-seq upon DNA transfection reveals transcriptomic changes. For technical inquiries, contact Ascent Research.
