The IFI16 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of the human A-549 lung adenocarcinoma epithelial cell line. This loss-of-function model disrupts the IFI16 gene, generating a heterogeneous cell pool with targeted gene ablation suitable for pooled functional studies. The polyclonal format provides a practical tool for investigating IFI16-dependent phenotypes without the need for monoclonal selection, enabling experiments that capture cellular heterogeneity.
The A-549 cell line, derived from the lung adenocarcinoma of a 58-year-old Caucasian male, serves as a widely used alveolar type II epithelial model. These cells retain characteristics of respiratory epithelium, including barrier function and drug-metabolizing enzyme activity, and are employed in pulmonary biology, toxicology, and cancer research. Their tumor origin also makes them a relevant substrate for studying oncogenic pathways and host?Ctumor immune interactions.
IFI16 (interferon gamma-inducible protein 16) is a cytosolic double-stranded DNA sensor that engages STING to activate TBK1 and IRF3, driving type I interferon production, while also stimulating NF-??B to induce pro-inflammatory cytokines. IFI16 additionally interacts with p53 to reinforce apoptosis and cellular senescence, functioning as a tumor suppressor. Key pathway members include cGAS, ASC, and caspase-1, with regulation by interferon-??, viral dsDNA, and DNA damage. Disruption of IFI16 in A-549 cells abolishes STING-dependent innate immune signaling and impairs p53-mediated apoptosis, as outlined in the mechanistic summary.
In the A-549 context, IFI16 knockout creates a system deficient in cytosolic DNA sensing, enabling dissection of the cGAS-STING pathway in tumor cell-autonomous immune responses. The loss also attenuates p53-dependent apoptosis and senescence, providing a platform to study how escape from IFI16-mediated tumor suppression might facilitate immune evasion or resistance to genotoxic therapy. This model thus allows investigation of lung adenocarcinoma cell responses to viral mimetics, synthetic DNA ligands, or cytosolic self-DNA in the absence of a key innate sensor.
This polyclonal knockout supports diverse applications, including innate immunity studies, herpes simplex virus-1 infection models, and cGAS-STING pathway analysis. It is suitable for cytokine profiling by ELISA, western blotting for phospho-STING/TBK1/IRF3, RT-qPCR of IFNB1 and interferon-stimulated genes, and flow cytometric apoptosis assays. Additional uses include co-immunoprecipitation of STING complexes, senescence-associated ??-galactosidase staining, and RNA-seq. For drug discovery, the cells can be employed in screens targeting DNA-sensing pathways or STING agonists. Researchers seeking a robust tool for investigating DNA-driven innate immunity in a lung tumor background are encouraged to contact Ascent Research for further information.
