The IFI27 Knockout SK-HEP-1 Polyclonal Cells product comprises a polyclonal population of SK-HEP-1 cells that have undergone CRISPR/Cas9-mediated gene disruption at the IFI27 locus. This heterogeneous knockout pool provides a loss-of-function model for studying interferon-induced mitochondrial apoptosis and innate immune signaling in a liver adenocarcinoma context. The polyclonal format retains cellular diversity and is well-suited for functional screening and pathway analysis without the clonal selection artifacts often associated with single-cell-derived knockout lines.
SK-HEP-1 is a human hepatic adenocarcinoma cell line derived from ascites of a 52-year-old male patient. Displaying both epithelial and endothelial markers, these adherent cells are tumorigenic and widely employed to investigate the molecular mechanisms of hepatocarcinogenesis, tumor metastasis, and therapeutic resistance. Their dual marker expression provides a unique platform for exploring apoptosis regulation and interferon responsiveness in a cellular background that recapitulates certain aspects of tumor stroma interactions.
IFI27 is an interferon-stimulated gene whose expression is rapidly induced by type I interferons (IFN-??/??) through the canonical JAK-STAT pathway. Binding of IFN-??/?? to the IFNAR1/IFNAR2 receptor activates JAK1 and TYK2, which phosphorylate STAT1 and STAT2. Phosphorylated STAT1/STAT2 associates with IRF9 to form the ISGF3 complex, which translocates to the nucleus and binds ISRE elements, driving IFI27 transcription. Additional induction can occur via IRF3 and IRF7 downstream of viral RNA sensors RIG-I and MDA5. The IFI27 protein localizes to the mitochondrial outer membrane where it engages anti-apoptotic Bcl-2 and Bcl-xL, disrupts mitochondrial membrane potential, and promotes the release of cytochrome c. This initiates apoptosome formation, caspase-9 activation, and subsequent cleavage of executioner caspase-3, culminating in mitochondrial-mediated cell death.
CRISPR-mediated knockout of IFI27 in the SK-HEP-1 cell line creates a valuable model for deciphering the role of interferon-induced apoptosis specifically in liver cancer. Given SK-HEP-1??s expression of both epithelial and endothelial markers, IFI27 loss-of-function can reveal context-dependent survival mechanisms that may contribute to hepatocellular carcinoma progression or resistance to interferon-based therapies. This polyclonal knockout pool also facilitates the identification of synthetic lethal interactions and the evaluation of small-molecule modulators targeting Bcl-2 family proteins or upstream interferon signaling components.
Researchers can employ this model in a variety of apoptosis and signaling assays, including Annexin V and JC-1 staining to quantify apoptosis and mitochondrial depolarization, Western blotting and RT-qPCR to monitor IFI27 and downstream effector levels, co-immunoprecipitation to confirm Bcl-2/Bcl-xL interactions, and caspase activity measurements. The system is well-suited for drug response profiling, genetic modifier screens, and mechanistic studies of antiviral innate immunity in hepatic cells. For additional product information, technical support, or aliquoting options, please contact Ascent Research.
