TCR/B2M Knockout NFAT Luciferase Reporter Jurkat Cell Line
This cell line is a double knockout of TCR (T Cell Receptor) and B2M (Beta-2-Microglobulin). First, the TRAC (T-Cell Receptor Alpha Constant) and the TRBC1 (T-Cell Receptor Beta Constant 1) domains of the TCRα/β chains were genetically removed by CRISPR/Cas9 genome editing from NFAT Luciferase Reporter Jurkat cells to generate the TCR Knockout NFAT Luciferase Reporter Jurkat cell Line (BPS Bioscience #78556). These TCR knockout cells were used to generate a new cell line in which B2M was also genetically removed by CRISPR/Cas9 genome editing.
Expression of the firefly luciferase gene is driven by NFAT response elements located upstream of the minimal TATA promoter. Activation of the NFAT signaling pathway in these cells can be monitored by measuring luciferase activity.
Purchase of this cell line is for research purposes only; commercial use requires a separate license. View the full terms and conditions.
Media Required for Cell Culture
| Name | Ordering Information |
| Thaw Medium 2 | BPS Bioscience #60184 |
| Growth Medium 2B | BPS Bioscience #79530 |
Materials Required for Cellular Assay
| Name | Ordering Information |
| Anti-CD3 Agonist Antibody | BPS Bioscience #71274 |
| NFAT Luciferase Jurkat Recombinant Cell Line | BPS Bioscience #60621 |
| ONE-Step™ Luciferase Assay System | BPS Bioscience #60690 |
| 96-well tissue culture-treated white, clear-bottom plate | |
| Luminometer |
Each vial contains 2 x 106 cells in 1 ml of cell freezing medium (BPS Bioscience #79796)
The cell line has been screened to confirm the absence of Mycoplasma species
The TCR (T Cell Receptor) is found on the surface of T-cells and is responsible for recognizing antigens bound to MHC (Major Histocompatibility Complex) molecules. Stimulation of the TCR results in activation of downstream NFAT signaling. NFAT (Nuclear factor of activated T-cells) is a family of transcription factors that has an important function in immune responses, for example by inducing the expression of various cytokines (such as interleukin-2 to 4, and TNF-alpha) in T cells. NFAT is regulated by Ca2+ and the Ca2+/calmodulin-dependent serine phosphatase, calcineurin.
Beta-2-Microglobulin is a required component of MHC class 1 molecules, which present peptide fragments from within the cell to cytotoxic T cells as part of the adaptive immune system. The protein forms amyloid fibrils in some pathological conditions. B2M plays an essential role both in governing MHC class I molecule stability and in promoting antigen binding and presenting the antigen to CD3/TCR complex of CD8+ T cells.
The knockout of both TCR and B2M will support the manufacture of universal CAR-T cells. The ablation of B2M or TCR prevents the elimination of allogeneic T cells that express foreign HLA-I molecules, and thereby enables the generation of CAR-T cells from allogeneic healthy donors T cells with higher persistence in vivo.
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