The IFI30 Knockout Jurkat Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat T lymphoblastoid line, engineered to disrupt the IFI30 gene. This loss-of-function model targets the lysosomal thiol reductase GILT, enabling population-level studies of antigen processing without clonal isolation.
The Jurkat parental line, a peripheral blood leukemia-derived T cell model (clone E6-1) with mutant p53, is widely employed to examine T lymphocyte signaling, proliferation, and apoptosis. Upon appropriate stimulation, Jurkat cells upregulate MHC class II antigen presentation components, providing a physiologically relevant backdrop for investigating the processing of protein antigens in a T cell environment.
GILT, encoded by IFI30, is transcriptionally induced by interferon gamma (IFNG) via the JAK1/JAK2-STAT1 pathway and acts as a critical lysosomal thiol reductase. Within the endocytic compartment, GILT reduces disulfide bonds of internalized antigens, facilitating their unfolding and proteolysis by cathepsins such as CTSS and CTSL. This reaction is essential for generating peptide ligands that associate with MHC class II heterodimers (HLA-DR, -DP, -DQ) in conjunction with the invariant chain CD74 and the peptide editor HLA-DM, ultimately enabling CD4+ T cell receptor recognition.
Knocking out IFI30 in Jurkat cells directly interrogates the redox-dependent steps governing MHC class II-restricted epitope supply. Because Jurkat cells can be driven to present exogenous antigens via surface MHC class II, the IFI30 disruption uniquely reveals how GILT deficiency impacts cathepsin-dependent antigen processing, peptide loading, and subsequent CD4+ T cell activation. This model holds particular relevance for dissecting immunodeficiencies and autoimmune disorders linked to faulty antigen handling and lysosomal redox imbalance.
Typical experimental applications include flow cytometric analysis of interferon gamma-induced MHC class II surface expression, ovalbumin-based antigen presentation assays measuring IL-2 secretion, and western blotting for key pathway components such as GILT, HLA-DR, and cathepsins. Additional uses encompass lysosomal enzymatic profiling and high-throughput screening of modulators targeting the antigen processing machinery. For further details or assistance, please contact Ascent Research.
