The CD47 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of Jurkat cells with targeted disruption of CD47. Generated by introducing Cas9 and a CD47-specific guide RNA, followed by enrichment for knockout cells without single-cell cloning, these cells retain polyclonal diversity, reducing clonal artifacts in functional studies and providing a loss-of-function model for CD47 in a T-cell context.
The host cell line, Jurkat, is a human T-lymphoblastoid line established from the peripheral blood of a 14-year-old patient with acute lymphoblastic leukemia. Jurkat cells are widely employed as a model system for T-cell receptor (TCR) signaling, apoptosis, and HIV infection. They express key T-cell markers, respond to TCR/CD3 stimulation, and secrete IL-2 upon activation. Their well-characterized signaling machinery and ease of genetic manipulation make them an ideal background for investigating immune checkpoint molecules such as CD47.
CD47 encodes a multi-pass transmembrane protein that functions as a ??don??t eat me?? signal by engaging signal regulatory protein alpha (SIRP??) on macrophages, thereby triggering phosphorylation of SHP-1/SHP-2 phosphatases and inhibiting phagocytosis. In T lymphocytes, CD47 also modulates integrin affinity and TCR signaling. CD47 expression is transcriptionally regulated by NF-??B, MYC, and HIF-1??, and is induced by inflammatory cytokines including TNF-?? and IFN-??. Its key interactions include binding to thrombospondin-1 (TSP-1) and integrins ??v??3 and ??IIb??3, as well as functional cross-talk with VEGFR2 and G??i proteins. Disruption of CD47 thus abrogates SIRP??-mediated inhibitory signals and alters integrin-dependent adhesion and T-cell activation.
In the Jurkat T-cell context, CD47 knockout provides a unique tool to dissect the role of CD47 in T-cell signaling independently of its anti-phagocytic function. Jurkat cells do not express macrophage-specific SIRP??, yet CD47 still modulates TCR-dependent activation and integrin activation. Thus, knocking out CD47 in Jurkat cells permits the study of T-cell-intrinsic functions of CD47, including its impact on NF-??B signalling, integrin affinity modulation, and interactions with TSP-1. This model is particularly relevant for understanding how CD47 influences T-cell immunity and immune evasion mechanisms in the tumor microenvironment.
Researchers can employ these polyclonal CD47 knockout Jurkat cells in a variety of experimental settings. Flow cytometry can confirm loss of CD47 surface expression, while phagocytosis assays can be performed by co-culturing with macrophages to measure antibody-dependent cellular phagocytosis. Western blotting can assess changes in SIRP?? phosphorylation and downstream SHP-1/SHP-2 activation. Additionally, T-cell activation assays, migration assays, and integrin activation assays enable functional evaluation of CD47??s role in T-cell adhesion and signaling. These cells are suitable for immune checkpoint research, cancer immunotherapy target validation, and mechanistic studies of CD47-SIRP?? interactions. For technical inquiries or to place an order, please contact Ascent Research.
