The IFI27 Knockout UM-UC-3 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the UM-UC-3 human bladder transitional cell carcinoma line, serving as a loss-of-function model for the IFI27 gene. This product consists of a heterogeneous pool of cells, each harboring distinct disruptions at the IFI27 locus, thereby enabling population-based functional studies without the constraints of clonal selection. The polyclonal format is particularly advantageous for experiments requiring robust statistical sampling, such as dose-response analyses of interferon treatment or apoptosis induction.
The parental UM-UC-3 cell line is derived from a high-grade human bladder transitional cell carcinoma and carries a mutant TP53 tumor suppressor gene, rendering it a widely recognized model for muscle-invasive bladder cancer. These cells display aggressive growth characteristics and invasive potential, making them a standard host for investigating oncogenic pathways, drug resistance mechanisms, and innate immune regulation in urothelial carcinoma.
IFI27 is an interferon-stimulated gene that encodes a mitochondrially localized protein critically involved in type I interferon-induced apoptosis. Mechanistically, IFI27 expression is driven by type I interferons (IFN-??/??) binding to the IFNAR1/IFNAR2 receptor complex, which activates the JAK1 and TYK2 kinases. This leads to phosphorylation and assembly of the ISGF3 transcription factor complex??comprising STAT1, STAT2, and IRF9??which then translocates to the nucleus and binds interferon-stimulated response elements (ISRE) in the IFI27 promoter. Once expressed, IFI27 translocates to mitochondria and physically interacts with the pro-apoptotic proteins BAX and BAK, facilitating mitochondrial outer membrane permeabilization. This process results in cytochrome c release and activation of the executioner caspases Caspase-3 and Caspase-7, culminating in apoptosis. IFI27 also contributes to antiviral immunity, and its function can be antagonized by viral proteins such as influenza NS1.
In bladder cancer, especially in the TP53-mutant background of UM-UC-3 cells, the intrinsic apoptotic machinery is frequently compromised. The IFI27 knockout enables precise dissection of the IFN-dependent apoptosis pathway, allowing researchers to determine whether IFI27 serves as a critical mitochondrial amplifier of cell death in carcinoma cells. This model is invaluable for studying immune evasion strategies employed by bladder tumors and for evaluating therapeutic approaches aimed at restoring or enhancing interferon-mediated cytotoxicity.
Key applications include Western blotting and RT-qPCR to verify IFI27 ablation, IFN-?? stimulation assays to interrogate JAK-STAT signaling, Annexin V staining for apoptosis detection, caspase-3/7 activity assays, and mitochondrial membrane potential measurements. The knockout cells are well-suited for paired experiments with wild-type UM-UC-3 controls to identify IFI27-specific responses during viral infection, cytokine challenge, or chemotherapeutic treatment. For further information, technical support, or to order, please contact Ascent Research.
