Background
The activation of T-cell receptors (TCRs) is a critical component of the adaptive immune response, driven by the interaction between TCRs and peptide-major histocompatibility complex (MHC) molecules on antigen-presenting cells (APCs). Understanding this interaction is pivotal for developing improved checkpoint and cell-based immunotherapies. To support this research, we have developed engineered cell lines and constructs to characterize antigen specificity and identify epitopes.
- The Jurkat TCR Knockout Cell Line (Jurkat TCRa/b-Knockout, CD8 Cells, Clone) was developed by CRISPR/Cas9-mediated knockout of the TRAC and TRBC genes in Jurkat E6.1 cells. The inactivation of TRAC/TRBC was confirmed via sequencing and negative staining with a PE-labeled anti-human TRBC1 antibody (Figure 1), which targets the TCR protein.
- The Jurkat TCR Knockout NFAT-GFP Reporter Cell Line (Jurkat TCRa/b-Knockout NFAT-tagGFP2 CD8, Clone) was transduced with GFP reporter construct expressing TCR: NFAT–mCMV–GFP–EF1–CD8 (Figure 2). Clonal selection was performed to isolate the cell line that showed the highest level of GFP expression—up to a 1,000-fold increase—after stimulation with PMA + ionomycin, indicating strong reporter activation.
- The K562 HLA-A-02:01 Cell line (Bleo or Puro, Polyclonal) was developed by engineering K562 cells to express the HLA-A-02:01 complex.
Applications:
- TCR Cloning and Evaluation:
Jurkat TCR-Reporter Cells engineered to express TCRs can be used to analyze the specificity and affinity of both native and engineered TCRs.
- Antigen Screening:
Libraries of specific peptides can be screened against TCR-expressing Jurkat TCR-reporter cells to identify those recognized by particular TCRs, as well as analysis of antibodies or small molecules that can mimic or inhibit TCR activation.
- T Cell Signaling Studies:
Jurkat TCR-Reporter Cells can be used to study T-cell pathway responses to tumor- or virus-specific antigens and to dissect the molecular mechanisms underlying T-cell activation
Please read the entire user manual before proceeding with your experiment.
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