The ARHGAP35 Knockout Jurkat Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal cell population derived from the Jurkat human T lymphocyte line, designed to disrupt the ARHGAP35 gene. This product provides a heterogeneous knockout pool suitable for loss-of-function studies, avoiding clonal selection artifacts. As a polyclonal population, it retains diverse genetic backgrounds with targeted ARHGAP35 disruption, enabling robust functional analysis of Rho GTPase signaling in T cells.
Jurkat cells are an immortalized human T-cell line originally isolated from the peripheral blood of a 14-year-old male with acute T-cell leukemia. This suspension-adapted line is a widely accepted model for investigating T-cell receptor (TCR) signaling, activation-induced proliferation, apoptosis, and leukemogenesis. Jurkat cells express key TCR complex components and downstream effectors, making them particularly suitable for dissecting signal transduction pathways governing immune cell function and malignant transformation.
ARHGAP35 encodes a Rho GTPase-activating protein (RhoGAP) that specifically inactivates RhoA, Rac1, and Cdc42 by accelerating their intrinsic GTP hydrolysis. This protein acts downstream of integrin engagement, receptor tyrosine kinases such as EGFR and PDGFR, and Src family kinases, including FAK. By terminating RhoA signaling, ARHGAP35 reduces actomyosin contractility and stress fiber formation, modulating actin dynamics through the ROCK-LIMK-cofilin axis. Additionally, ARHGAP35 interacts with p120RasGAP, cortactin, and filamin A, linking GTPase regulation to cytoskeletal scaffolding. Its activity ultimately influences SRF/MAL-dependent transcriptional programs associated with adhesion and proliferation.
In Jurkat T cells, ARHGAP35 functions as a critical regulator of immune synapse architecture and TCR signal transduction. Knockout of ARHGAP35 is expected to elevate RhoA-GTP levels, enhancing ROCK-mediated phosphorylation of LIMK and cofilin, thereby stabilizing F-actin. This cytoskeletal reconfiguration can alter immune synapse formation, T-cell activation marker expression (e.g., CD69, CD25), and migratory capacity. Given the leukemic origin of Jurkat cells, this model permits evaluation of how dysregulated Rho signaling contributes to malignant T-cell phenotypes, including unchecked proliferation and survival advantage.
Researchers can apply this knockout model to study T-cell activation dynamics, Rho GTPase contributions to leukemogenesis, and preclinical screening of Rho pathway inhibitors. Representative assays include RhoA G-LISA activation measurements, Western blot analysis of phospho-cofilin, flow cytometric assessment of activation markers, Transwell migration tests, phalloidin staining for F-actin, and immunofluorescence for immune synapse visualization. Additional applications encompass cell adhesion and CFSE proliferation assays to interrogate integrin-mediated functions and growth control. For further technical details, please contact Ascent Research.
