TCR Knockout NFAT-Luciferase-eGFP Reporter Jurkat Cell Line

TCR Knockout NFAT Luciferase-eGFP Reporter Jurkat Cell Line is a Jurkat cell line generated by using NFAT-Luciferase-eGFP Reporter Lentivirus (BPS Bioscience #78656-G) to transduce TCR Knockout Jurkat Cell Line (BPS Bioscience #78539). In this model, the TRAC (T-cell receptor alpha constant) and TRBC1 (T-cell receptor beta constant 1) regions of the TCR alpha/beta chains were deleted using CRISPR/Cas9 genome editing. The resulting Jurkat cells stably express a firefly luciferase reporter and an eGFP reporter, both under the control of NFAT response elements, enabling quantitative assessment of NFAT signaling activity. This cell line has been functionally validated and does not respond to anti-CD3 agonist antibodies, as opposed to the parental NFAT-Luciferase Reporter Jurkat Cell Line (BPS Bioscience #60621). Loss of expression of TCRalpha/beta in the TCR Knockout NFAT Luciferase-eGFP Reporter Jurkat Cell Line cells had been confirmed by flow cytometry and genomic sequencing. 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 (Nuclear factor of activated T-cells) signaling. NFAT 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. The TCR consists of a heterodimer of two different protein chains, of which the alpha (alpha) and beta (beta) chains are the predominant chains. CRISPR/Cas9 genome editing was used to remove the TRAC (T-Cell Receptor Alpha Constant) and TRBC1 (T-Cell Receptor Beta Constant 1) regions of the alpha and beta chains, resulting in loss of TCR expression. The removal of endogenous TCRs allows the expression of transgenic TCRs at higher levels and increases the efficiency of the edited T cells. Thus, knocking out the endogenous TCRs is becoming part of the T cell production workflow in clinical applications.