The IFI30 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human NCI-H1975 lung adenocarcinoma cell line. This product provides a heterogeneous pool of cells with targeted disruptions in the IFI30 gene, which encodes gamma-interferon-inducible lysosomal thiol reductase (GILT). The polyclonal format, generated via CRISPR/Cas9 technology, avoids single-cell clonal selection and provides a robust model for studying bulk antigen presentation and downstream immune responses.
The parental NCI-H1975 line is a well-characterized non-small cell lung cancer (NSCLC) model harboring EGFR L858R and T790M mutations, which drive oncogenic signaling and confer resistance to first-generation tyrosine kinase inhibitors. This genetic background is highly relevant for investigating tumor immune evasion and targeted therapy mechanisms in EGFR-mutant lung adenocarcinoma.
IFI30 encodes GILT, an endocytic thiol reductase essential for reducing disulfide bonds in internalized protein antigens. GILT activity facilitates antigen unfolding and subsequent proteolysis by cathepsins (including Cathepsin S and L) and legumain, enabling MHC class II-restricted peptide presentation. IFI30 expression is induced by interferon-gamma (IFNG) through the JAK1/2-STAT1 pathway and IRF1 transcription factor. GILT collaborates with CD74 and HLA-DM to load peptides onto MHC class II molecules, leading to T cell receptor engagement on CD4+ T cells and downstream cytokine production such as IFNG and interleukin-2. Knockout of IFI30 disrupts this pathway, impairing antigen processing and reducing surface MHC class II expression.
In the NCI-H1975 EGFR-mutant background, loss of GILT provides a model to study how redox-mediated antigen processing intersects with oncogenic signaling and immune recognition. This knockout system is particularly suited to examine whether impaired MHC class II presentation enhances tumor immune evasion, and how interferon signaling may be rewired in lung cancer cells to modulate T cell responses. Researchers can explore potential synergies with immune checkpoint blockade or adoptive T cell therapies.
Typical applications include T cell co-culture assays to measure antigen-specific activation by flow cytometry, antigen degradation studies to assess GILT-dependent processing, immunofluorescence for lysosomal markers, and evaluation of autophagy flux. Western blotting and RT-qPCR confirm IFI30 disruption, while surface MHC-II levels can be monitored under IFNG stimulation. The cells also support drug screening for immunomodulators. For technical inquiries, please contact Ascent Research.
