The IFI16 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the IFI16 gene has been disrupted to provide a high-quality loss-of-function model. This polyclonal pool is designed for researchers studying innate immune responses, DNA sensing, and inflammation, enabling robust and reproducible experimental outcomes in a well-characterized human cell background.
The host HAP1 cell line is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line, exhibiting an adherent, fibroblast-like morphology. With its near-haploid karyotype, HAP1 offers a simplified genetic landscape that reduces functional redundancy and facilitates straightforward gene-editing studies, making it particularly valuable for genetic screening and functional genomics. This unique ploidy ensures that disruption of a single allele is sufficient to ablate gene function, providing a clean background for investigating gene function without confounding diploid buffering.
IFI16 functions as a critical sensor of cytosolic double-stranded DNA, acting upstream of the cGAS-STING pathway to initiate antiviral and inflammatory responses. Upon DNA recognition, IFI16 engages STING, leading to the activation of TBK1 and subsequent phosphorylation of IRF3, which drives the expression of type I interferons such as IFN-?? and interferon-stimulated genes (ISGs). In addition to STING-dependent signaling, IFI16 can assemble an inflammasome complex with ASC and caspase-1, promoting the maturation and secretion of IL-1?? and inducing pyroptosis. Furthermore, IFI16 interacts with p53 and BRCA1, enhancing p53-mediated transcriptional activity to induce cell cycle arrest via p21 and apoptosis. These multifaceted roles position IFI16 as a pivotal node linking DNA sensing to innate immunity, cell death, and tumor suppression.
Disrupting IFI16 in the near-haploid HAP1 background creates a potent tool for dissecting these signaling networks without the complication of allelic variation. The knockout cells enable precise analysis of IFI16-dependent pathways, distinguishing STING-mediated interferon induction from inflammasome-driven inflammatory responses. This model is especially advantageous for interrogating the interplay between DNA damage responses and innate immunity, as the p53-IFI16 axis can be studied in isolation. Researchers can thus delineate the contributions of IFI16 to cell fate decisions, including senescence and programmed cell death, in a genetically tractable system.
These IFI16 Knockout HAP1 Polyclonal Cells are suitable for a wide array of experimental applications, including investigation of DNA sensing mechanisms, STING signaling cascades, inflammasome activation, tumor suppressor pathways, viral pathogenesis, and autoimmune disease research. Representative assay strategies include western blotting for phosphorylated IRF3 or STING, RT-qPCR for IFN-?? and ISG expression, ELISA for IFN-?? and IL-1?? secretion, immunofluorescence to assess IFI16 subcellular localization, co-immunoprecipitation to probe IFI16-STING interactions, luciferase-based IFN-?? promoter reporter assays, and flow cytometry to measure apoptosis. DNA stimulation assays using synthetic ligands or viral infection models further enable functional readouts of pathway integrity. For additional details and ordering information, please contact Ascent Research.
