The ITPKC Knockout Jurkat Polyclonal Cells are a polyclonal population of Jurkat T-lymphoblastoid cells engineered by CRISPR/Cas9-mediated disruption of the ITPKC gene. This product provides a heterogeneous knockout model, with each cell carrying a distinct gene-editing event, enabling robust loss-of-function analysis without clonal bias. As a polyclonal knockout pool, these cells are well-suited for experiments requiring rapid generation of knockout phenotypes and for applications where the complexity of polyclonal editing mirrors physiological heterogeneity. The cells are validated to express the Jurkat surface markers and demonstrate disrupted ITPKC expression at the protein level.
The host cell line, Jurkat, clone E6-1, is an immortalized human T-lymphocyte line derived from the peripheral blood of a 14-year-old male with acute lymphoblastic leukemia (ALL). Jurkat cells are extensively used to dissect T cell receptor (TCR) signaling, activation, and apoptosis, owing to their well-characterized signaling pathways and responsiveness to stimuli such as anti-CD3/CD28 antibodies. They serve as a model for acute T cell leukemia and for studying the molecular mechanisms of T cell activation and immune regulation, providing a relevant cellular context for exploring the function of ITPKC.
ITPKC encodes inositol 1,4,5-trisphosphate 3-kinase C, which phosphorylates the second messenger inositol 1,4,5-trisphosphate (IP3) to inositol 1,3,4,5-tetrakisphosphate (IP4). This conversion limits IP3-mediated calcium release from the endoplasmic reticulum by reducing the pool of IP3 available to activate the IP3 receptor. In T cells, upon TCR engagement, PLC??1 generates IP3, triggering Ca2+ release, which activates calmodulin and calcineurin, leading to dephosphorylation and nuclear translocation of NFAT transcription factors. ITPKC, activated by Ca2+/calmodulin, negatively regulates this cascade by shunting IP3 towards IP4, thereby dampening calcium flux and subsequent NFAT-dependent transcription of genes such as IL-2. ITPKC thus functions as a critical rheostat in the TCR signaling pathway, modulating the intensity and duration of calcium signals.
In the Jurkat cell background, loss of ITPKC function is predicted to potentiate TCR-induced calcium mobilization and NFAT activation, leading to enhanced IL-2 expression and a hyperresponsive phenotype. This makes the ITPKC knockout model particularly relevant for investigating the dysregulated T cell activation observed in Kawasaki disease, where ITPKC polymorphisms are associated with susceptibility and coronary artery lesion formation. Moreover, given the leukemic origin of Jurkat cells, this knockout model provides a platform for examining the role of calcium and NFAT signaling in T-cell acute lymphoblastic leukemia proliferation and survival, potentially revealing therapeutic vulnerabilities.
Researchers can employ these ITPKC knockout cells to dissect the calcium-NFAT signaling axis using assays such as NFAT nuclear translocation immunofluorescence, calcium flux measurements with Fluo-4, and NFAT-luciferase reporter assays. Additional methods include western blotting for phosphorylated ITPKC, RT-qPCR for IL-2 and NFAT targets, and ELISA for IL-2 secretion. The knockout cells are suitable for drug screening to identify modulators of T cell activation and for comparative studies with wild-type Jurkat cells. For further technical details or ordering information, please contact Ascent Research.
