ARFIP2 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T lymphocyte line. This loss-of-function model provides a system to investigate the ADP-ribosylation factor interacting protein 2 (ARFIP2) gene in an immune cell context, enabling studies of its roles in membrane trafficking and actin cytoskeleton regulation.
Jurkat cells are an immortalized human T lymphocyte line originally isolated from a 14-year-old male with acute T cell leukemia. Widely employed as a model for T cell receptor (TCR) signaling, leukemia biology, and immune synapse formation, Jurkat cells exhibit robust activation upon TCR/CD3 stimulation, making them suitable for dissecting signaling pathways relevant to adaptive immunity.
ARFIP2 encodes a scaffold protein that directly interacts with GTP-bound ARF1, ARF5, ARF6, and Rac1 GTPases, linking vesicular trafficking to actin dynamics. Downstream of TCR/CD3-mediated Rac1 activation, ARFIP2 promotes actin polymerization and membrane ruffling through effectors including the WAVE and WASP proteins, the Arp2/3 complex, and the LIMK-cofilin axis. It also facilitates Golgi-derived vesicle formation necessary for NFAT translocation, and interacts with PIP5K to modulate phosphoinositide metabolism, thereby integrating membrane transport with cytoskeletal remodeling.
In Jurkat cells, ARFIP2 disruption impairs TCR-induced actin reorganization, immune synapse formation, and subsequent NFAT activation, resulting in attenuated IL-2 secretion and CD69 expression. This knockout model thus enables precise analysis of how ARFIP2-dependent trafficking coordinates T cell activation. As ARFIP2 has been implicated in cancer metastasis, prostate cancer, hepatocellular carcinoma, and neurodegenerative disorders, these cells also provide a platform for investigating broader pathological mechanisms.
Researchers typically confirm target gene disruption by RT-qPCR and Western blotting, followed by functional analyses such as flow cytometry for activation markers (CD69), ELISA for IL-2 secretion, and immunofluorescence microscopy to assess actin remodeling. Co-immunoprecipitation and Western blotting facilitate examination of protein-protein interactions, and cell migration or apoptosis assays enable study of ARFIP2’s role in leukemia cell invasiveness and survival. The polyclonal knockout population is well-suited for drug discovery efforts targeting the ARF/Rac signaling axis or screening for modulators of immune synapse formation. For technical inquiries or support, please contact Ascent Research.
