The IFI16 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the IFI16 gene has been disrupted to establish a loss-of-function model. Derived from the NCI-H1975 non-small cell lung adenocarcinoma line, this heterogeneous pool of edited cells avoids clonal selection artifacts and better reflects the genetic diversity of tumor cell populations while abrogating IFI16 protein expression.
The parental NCI-H1975 line is a well-characterized model of human lung adenocarcinoma, harboring an activating EGFR L858R mutation and a TP53 loss-of-function mutation. These epithelial cells are widely used to study oncogenic signaling, drug resistance, and the interplay between EGFR-driven pathways and innate immune responses in non-small cell lung cancer.
IFI16 is a cytosolic double-stranded DNA sensor that activates innate immunity by engaging STING, which triggers TBK1-mediated phosphorylation of IRF3, leading to transcriptional induction of type I interferons such as IFNB1. IFI16 also forms an inflammasome complex with ASC to activate caspase-1 and promotes p53-dependent apoptosis and senescence through interactions with BRCA1 and p53. This knockout disrupts the cGAS-STING-IFI16-TBK1-IRF3 axis and the DNA damage response involving p53.
In the NCI-H1975 background, IFI16 loss enables dissection of DNA sensing and immune evasion mechanisms in the context of defective p53 and active EGFR signaling. Researchers can investigate how the absence of IFI16 alters STING-dependent interferon production, inflammasome activity, and apoptosis independent of p53, providing insights into innate immune regulation in lung adenocarcinoma and potential therapeutic vulnerabilities.
Applications include western blotting for key signaling proteins (e.g., TBK1, IRF3, STAT1), RT-qPCR for interferon-stimulated genes, immunofluorescence to assess IFI16 localization, co-immunoprecipitation with STING or ASC, and reporter assays for interferon-beta promoter activity. Apoptosis assays (Annexin V staining) and DNA stimulation experiments can define IFI16??s role in cell death and immune activation. This tool is invaluable for innate immunity, viral infection, cancer immunology, and drug target validation studies. For further information, please contact Ascent Research.
