The IFI30 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from SK-HEP-1 human liver adenocarcinoma cells, harboring disruption of IFI30. This heterogeneous pool serves as a loss-of-function model to dissect the roles of IFI30, a lysosomal thiol reductase, in antigen processing and immune modulation. The polyclonal format captures diverse editing events, enabling studies that reflect a range of knockout phenotypes while avoiding clonal artifacts.
SK-HEP-1 cells were isolated from the ascites of a 52-year-old male hepatic adenocarcinoma patient and exhibit a mixed endothelial?Cepithelial phenotype, expressing markers such as CD34, Factor VIII-related antigen, and epithelial determinants. Their aneuploid karyotype and metastatic nature make them a robust model for liver cancer biology and tumor angiogenesis. Targeting IFI30 in this background provides a relevant platform for studying lysosomal function and immune evasion in an epithelial tumor context.
IFI30 encodes a lysosomal oxidoreductase that reduces disulfide bonds of endocytosed antigens, enabling proteolytic processing and loading onto MHC class II molecules. IFI30 expression is induced by interferon gamma (IFNG) via STAT1 and regulated by NF-??B and LPS. Within the lysosome, IFI30 interacts with CD74, cathepsin S, Hsc70, and LAMP-2 to facilitate antigen unfolding. This yields reduced peptides that are loaded onto MHC class II with the help of HLA-DM, leading to CD4+ T cell activation. Thus, IFI30 links lysosomal redox regulation to adaptive immunity.
Knocking out IFI30 in SK-HEP-1 cells allows investigation of how tumor-intrinsic antigen processing affects MHC class II presentation and immune surveillance. The endothelial-like characteristics of this line enable exploration of crosstalk between thiol reductase activity and angiogenic signaling. IFI30 loss impairs antigenic peptide generation, potentially reducing MHC class II-restricted T cell recognition and facilitating immune escape. This model is also relevant for lysosomal storage disorders and examining redox imbalance in malignant hepatocytes, with implications for autoimmunity and cancer immunotherapy.
This polyclonal IFI30 knockout model supports diverse applications: Western blotting and RT-qPCR for gene disruption validation, immunofluorescence and lysosomal fractionation for subcellular studies, and flow cytometry for MHC class II expression upon interferon gamma stimulation. Antigen presentation assays and T cell co-cultures can measure functional immune outcomes, while lysosomal biochemistry and stress response studies are also feasible. For further information, please contact Ascent Research.
