ARHGDIB Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat T lymphocyte cell line. This product provides a heterogeneous pool of cells with targeted disruption of the ARHGDIB gene, encoding Rho GDP dissociation inhibitor beta (RhoGDI2). The polyclonal format ensures a spectrum of knockout efficiencies across the population, enabling researchers to investigate gene dosage effects on T cell physiology. Unlike clonal lines, this population retains biological variability while maintaining robust loss-of-function of RhoGDI2.
Jurkat is an immortalized human T lymphocyte line established from the peripheral blood of a 14-year-old male with acute T cell leukemia. These suspension lymphoblast cells are widely used as a model for T cell receptor (TCR) signaling, apoptosis, and leukemia biology. Their genetic tractability and well-characterized signaling pathways make them ideal for studying cytoskeletal dynamics and immune cell function. The ARHGDIB knockout in this context allows dissection of RhoGDI2’s role in T cell-specific processes.
ARHGDIB encodes RhoGDI2, a pivotal regulator of the Rho GTPase cycle. RhoGDI2 sequesters Rho family GTPases??RhoA, Rac1, and Cdc42??in the cytosol, inhibiting nucleotide exchange and membrane association. This inhibition is relieved by signals from Src family kinases, protein kinase C, and ERM proteins (Ezrin, Radixin, Moesin), triggering GTPase activation and downstream actin polymerization, cell adhesion, and migration. Knockout of ARHGDIB disrupts this control, causing constitutive activation of Rho GTPases and altered signaling via integrin adhesion, chemokine-directed migration, and NF-??B transcription. Interacting partners include ARHGDIA, and downstream targets encompass focal adhesion kinase (FAK) and serum response factor (SRF). The polyclonal knockout population exhibits variable RhoGDI2 depletion, enabling study of concentration-dependent regulation of these cascades.
In Jurkat T cells, ARHGDIB knockout profoundly impacts immune function. Enhanced Rho GTPase activity drives cytoskeletal reorganization, altering cell morphology, adhesion, and transendothelial migration??critical for T cell trafficking and surveillance. This model is relevant for primary immunodeficiencies, T cell lymphomas, and other cancers where RhoGDI2 is implicated. It also facilitates study of RhoGDI2’s role in apoptosis and NF-??B signaling, pathways dysregulated in leukemia and inflammatory disorders. The polyclonal nature permits assessment of how varying RhoGDI2 levels affect these processes, mimicking heterozygous loss-of-function scenarios.
Researchers can use this knockout population for diverse functional studies, including dissecting TCR signaling, analyzing immune cell migration via Transwell assays, and measuring actin cytoskeleton changes by flow cytometry or immunofluorescence. The cells are suitable for G-LISA and pull-down assays to quantify active RhoA, Rac1, and Cdc42, and Western blotting to assess ARHGDIB ablation and downstream signaling. Phospho-kinase arrays and RNA-seq can further map pathway perturbations. This model is valuable for drug screening targeting Rho GTPase signaling in cancer and inflammation. For additional information or custom requests, please contact Ascent Research.
