The ARFGEF1 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population, designed to abolish expression of the ARFGEF1 (BIG1) gene in the Jurkat human T lymphocyte line. This mixed population provides a robust loss-of-function model for investigating the roles of ARFGEF1 in vesicular trafficking and signaling without requiring clonal isolation. The cells are suitable for a wide range of biochemical and cell-based assays in an immortalized T-cell context.
The Jurkat cell line is an immortalized human T lymphocyte line originally derived from the peripheral blood of a 14-year-old male with T-cell acute lymphoblastic leukemia. Suspension-adapted lymphoblasts, Jurkat cells are a widely accepted model for studying T-cell receptor (TCR) signal transduction, cytokine secretion, and apoptosis pathways. Their rapid growth and genetic tractability make them ideal for CRISPR editing and functional genomics.
ARFGEF1 encodes Brefeldin A-Inhibited Guanine nucleotide-exchange Protein 1 (BIG1), a critical activator of ADP-ribosylation factor 1 (ARF1). As a guanine nucleotide exchange factor (GEF), ARFGEF1 catalyzes GDP/GTP exchange on ARF1, enabling recruitment of COPI and clathrin adaptor complexes, including AP-1, to Golgi and endosomal membranes. ARFGEF1 is regulated by phosphatidylinositol 4,5-bisphosphate (PIP2) binding and phosphorylation by protein kinase A (PKA), and its activity is sensitive to brefeldin A treatment. It interacts with ARF1, ARFGEF2, FKBP13, the PKA regulatory subunit RI??, and myosin IIA. Downstream, ARF1-GTP promotes COPI coat assembly for retrograde Golgi-to-ER transport, AP-1 and GGA adaptor recruitment for TGN-to-endosome sorting, and phosphatidylinositol 4-phosphate 5-kinase (PI4P5K) activation to generate PIP2, thereby coordinating Golgi integrity, endosomal recycling, integrin trafficking, and actin cytoskeleton remodeling.
In Jurkat T lymphocytes, ARFGEF1-dependent ARF1 activation is implicated in regulating TCR recycling, integrin trafficking, and cytokine secretion pathways. Disruption of ARFGEF1 in this cell line provides a physiologically relevant model to dissect how ARF1-GTPase signaling influences immune synapse formation, Golgi stress responses, and secretory autophagy, processes that are frequently dysregulated in T-cell leukemia. The polyclonal knockout population allows assessment of heterogeneous knockout effects while maintaining the leukemic background critical for cancer biology studies.
This product is suitable for a broad spectrum of experimental applications, including western blot analysis of ARFGEF1 and ARF1-GTP levels, immunofluorescence staining of Golgi markers such as GM130 to examine organelle morphology, and flow cytometry-based assays for cell surface receptor recycling. Functional studies may encompass co-immunoprecipitation of ARF1 complexes, RT-qPCR profiling of downstream transcriptional targets, migration and invasion assays, and brefeldin A sensitivity testing. The cells are particularly valuable for investigating the molecular mechanisms of periventricular nodular heterotopia, breast and colorectal cancer metastasis, and T-cell leukemogenesis. For further technical details or ordering inquiries, please contact Ascent Research.
