The DLG1 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T lymphocyte line, designed to disrupt the DLG1 gene (Discs Large MAGUK Scaffold Protein 1) in Homo sapiens. This polyclonal pool offers a genetically heterogeneous loss-of-function model, generated by CRISPR/Cas9-mediated gene disruption without clonal isolation, enabling robust studies of DLG1-dependent signaling networks in a polyclonal context. The cells are provided as a ready-to-use knockout tool for investigating scaffold protein functions in T-cell receptor (TCR) signaling and associated pathways.
Jurkat cells are an immortalized T-cell line originally established from a patient with acute lymphoblastic leukemia (ALL). They serve as a widely employed model system for dissecting TCR-mediated signal transduction, T-cell activation, and leukemogenesis. Their well-characterized signaling machinery and ease of manipulation make them an ideal host for gene-editing approaches, including CRISPR/Cas9-based knockout strategies. Notably, Jurkat cells recapitulate key aspects of early T-cell activation events, including downstream phosphorylation cascades and cytokine production, thereby providing a physiologically relevant background for studying the impact of DLG1 disruption.
DLG1 encodes a membrane-associated guanylate kinase (MAGUK) scaffold protein that organizes supramolecular signaling complexes at tight junctions and the immunological synapse. It is regulated by upstream stimuli such as TCR engagement, CD28 co-stimulation, and phosphorylation by CaMKII, CDK5, and Src family kinases (Lck, Fyn). DLG1 directly interacts with Lck, ZAP-70, APC, AMPK, CASK, LIN7, MPP7, actin, GKAP, and ERBB2, and scaffolds critical TCR pathway components including the TCR-CD3 complex, Lck, ZAP-70, LAT, and PLC??1. Downstream, DLG1 modulates targets such as p38 MAPK, NFAT, YAP, and AMPK, and controls actin cytoskeleton reorganization. Its function bridges TCR proximal signaling to the Hippo pathway via MST1/2?CLATS1/2?CYAP axis, influencing cell proliferation and polarity. Mechanistically, DLG1 facilitates efficient TCR signal transduction by anchoring Lck in proximity to ZAP-70, thereby enabling phosphorylation of downstream adaptors and effectors.
In the Jurkat T-cell context, DLG1 knockout disrupts the spatial organization of signaling molecules at the immunological synapse, leading to impaired TCR-induced activation. Loss of DLG1 is expected to reduce Lck-mediated ZAP-70 phosphorylation, attenuate NFAT and AP-1 transcriptional activity, and diminish IL-2 production, as assessed by luciferase reporters and ELISA. Additionally, dysregulation of the Hippo pathway may alter YAP-dependent proliferative responses, linking DLG1 function to cell growth control. This model thus provides a powerful system to dissect how scaffold protein-mediated complex assembly governs T-cell activation thresholds, synapse stability, and crosstalk with tumor suppressor pathways. Because Jurkat cells are of leukemic origin, the knockout also serves as a platform to study how DLG1 loss influences malignant T-cell behavior, including altered polarity and unchecked proliferation.
The DLG1 Knockout Jurkat Polyclonal Cells are ideally suited for a range of experimental applications, including validation of DLG1 loss by Western blotting, flow cytometric analysis of activation markers (CD69, CD25), phospho-flow detection of pZAP-70 and pLck, and co-immunoprecipitation of Lck-DLG1 complexes. They enable functional readouts using NFAT/AP-1 luciferase reporters, IL-2 ELISA, and CFSE-based proliferation assays. Researchers can employ this polyclonal knockout population to investigate TCR signal transduction mechanisms, immunological synapse assembly, Hippo pathway regulation, cancer cell polarity, and drug target validation. For further information, please contact Ascent Research.
