The HLA-C Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited population of HEK293T cells harboring targeted disruption of the HLA-C gene. Supplied as polyclonal cells, this reagent avoids clonal selection, enabling direct use in functional studies. The knockout eliminates HLA-C-mediated antigen presentation and NK cell inhibitory signaling, offering a clean genetic model to explore immune recognition mechanisms. Researchers can immediately compare this polyclonal knockout population with wild-type HEK293T cells in a range of immunological assays, ensuring experimental consistency.
HEK293T cells originate from human embryonic kidney epithelial cells immortalized by adenovirus 5 DNA. They constitutively express the SV40 large T antigen, facilitating high-yield transfection and viral production. This cell line exhibits robust growth and is widely adopted for protein expression, lentivirus packaging, and functional genomics. Although of epithelial origin, HEK293T cells retain endogenous expression of many immune-pathway components, making them a useful platform for dissecting cell-autonomous immune functions without the confounding variables of lymphoid lineage.
HLA-C is a classical MHC class I molecule that presents intracellular peptides to CD8+ T cells. It also functions as an inhibitory ligand for KIRs (e.g., KIR2DL1, KIR2DL2/3) on NK cells, preventing cytotoxic attack. The HLA-C?Cpeptide complex assembly involves B2M, TAP1/2, TAPBP, CALR, and PDIA3. Transcription is upregulated by IFN-?? via IRF1 and the master MHC class I transactivator NLRC5, and by TNF-?? through NF-??B. Thus, HLA-C sits at the intersection of innate and adaptive immunity, translating inflammatory signals into immune surveillance and self-tolerance.
Knockout of HLA-C in HEK293T cells abolishes surface MHC-I expression, removing the ligand for inhibitory KIRs and rendering cells sensitive to NK cell cytotoxicity. This phenotype isolated in an epithelial context enables precise analysis of HLA-C??s role without overlap from lymphocyte-specific regulators. The polyclonal nature of the edited population mirrors the genetic diversity found in primary tissues, increasing translational relevance for tumor immunology and transplantation biology. Consequently, these cells are a powerful tool to study mechanisms of immune evasion and allogeneic cell rejection.
Typical applications include measuring CD8+ T cell activation via IFN-?? ELISpot and flow cytometry for MHC-I after peptide loading, or assaying NK cell degranulation and cytotoxicity in co-culture experiments. The knockout background facilitates reconstitution studies of viral immune evasion proteins that target HLA-C trafficking or expression. In allogeneic cell therapy research, these cells permit evaluation of HLA-C deletion as a strategy to modulate NK-mediated rejection. For technical inquiries or protocol assistance, please contact Ascent Research.
