ZsGreen Knock-in TRAC Knock-out Jurkat Cell Line

ZsGreen Knock-in TRAC Knock-out Jurkat Cell Line is a Jurkat knock-in cell line that expresses ZsGreen (Zoanthus sp. green fluorescent protein) in the TRAC (T-Cell Receptor Alpha Constant) locus. As a result, the endogenous TRAC is knocked out and no longer expressed. This cell line was generated by electroporation with the Cas9 enzyme (Streptococcus pyogenes) and sgRNA (single guide RNA) targeting TRAC (BPS Bioscience #82394), along with transduction with an AAV HDR (Homologous Recombination template) featuring ZsGreen driven by an EF1a promoter. This cell line has been validated by flow cytometry, PCR (polymerase chain reaction), and sequencing. The T Cell Receptor (TCR) 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 IL-2/IL-3/IL-4 and TNF (tumor necrosis factor)-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. Removal of the TRAC (T-Cell Receptor Alpha Constant) region of the alpha chains results 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.