The ECE1 Knockout Jurkat Polyclonal Cells are a heterogeneous population of Jurkat T lymphocytes subjected to CRISPR/Cas9-mediated disruption of the ECE1 gene, which encodes endothelin-converting enzyme 1. This product serves as a loss-of-function model for investigating ECE1-dependent proteolytic processing and downstream signaling cascades in an immune cell context. As a polyclonal knockout pool, it reflects diverse editing events across the cell population, providing a system-level tool for functional genomics and drug discovery applications without the bias of a single clone.
The parental Jurkat cell line is an immortalized human T lymphocyte line derived from acute lymphoblastic leukemia. Widely utilized for studying T cell receptor signaling, apoptosis, and HIV infection, Jurkat cells offer a robust and well-characterized platform for interrogating molecular mechanisms in adaptive immunity. Their rapid growth and ease of genetic manipulation make them ideal for generating knockout models to assess gene function in T cell biology.
ECE1 is a membrane-bound metalloprotease that catalyzes the conversion of inactive big endothelin-1 to the potent vasoactive peptide endothelin-1, which engages ETA and ETB receptors to activate multiple signaling pathways, including Gq/11-mediated PLC?? stimulation, calcium mobilization, and PKC/ERK cascades. In addition, ECE1 processes bradykinin and substance P, linking it to inflammation and pain signaling. Within Jurkat T cells, ECE1 expression is regulated by factors such as TGF-??1, TNF-??, and HIF-1??, and its enzymatic activity contributes to autocrine/paracrine endothelin signaling that can enhance MAPK activation and NF-??B/AP-1 transcription factor responses. Downstream targets include ERK1/2, p38 MAPK, c-Fos, and c-Jun, while interacting proteins such as neprilysin and ADAM17 modulate its function.
In the Jurkat T lymphocyte model, ECE1 knockout disrupts endothelin-1 maturation, potentially attenuating endothelin-driven calcium signaling and MAPK pathway activation, which are critical for T cell activation, cytokine production, and proliferation. This loss-of-function model enables dissection of ECE1’s role in immune cell signaling beyond its classical cardiovascular functions. Given that endothelin-1 has been implicated in T cell migration and adhesion, the knockout cells provide a valuable tool for exploring how endothelin signaling intersects with adaptive immunity, including potential effects on integrin activation and chemotaxis. Moreover, because Jurkat cells are leukemic in origin, the model can be applied to study the relevance of ECE1 in hematologic malignancies and inflammation-associated cancer biology.
Research applications include investigating endothelin signaling in T cell biology, evaluating ECE1 function in immune cell activation and inflammation, screening for ECE1 inhibitors, and functional genomics of metalloproteases in leukemia. Representative assays well suited for this model are Western blotting and ELISA to assess endothelin-1 levels, RT-qPCR for transcript analysis, calcium mobilization assays to measure proximal signaling, phospho-ERK flow cytometry for MAPK activation, cytokine bead arrays for secretion profiling, and transwell migration assays to evaluate chemotaxis. The polyclonal knockout population allows robust, scalable experiments without clonal selection bias. For further information, please contact Ascent Research.
