ARHGEF40 Knockout Jurkat Polyclonal Cells constitute a ready-to-use CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the ARHGEF40 gene. This product provides a genetically heterogeneous pool of Jurkat cells harboring targeted disruptions in the ARHGEF40 locus, enabling investigation of gene function without the clonal selection biases associated with single-cell-derived knockout lines. The polyclonal format preserves the diversity of editing outcomes across the population, offering a robust and scalable model for studying ARHGEF40-dependent phenotypes in a human T lymphocyte background.
The host Jurkat cell line is an immortalized human acute T cell leukemia-derived line widely employed as a model for T cell receptor (TCR) signaling, cytokine production, and immune response mechanisms. Jurkat cells exhibit key characteristics of activated T lymphocytes, including the ability to form immunological synapses and produce interleukin-2 upon stimulation. Their genetic tractability and well-characterized signaling networks make them an ideal chassis for dissecting the molecular underpinnings of T cell activation and migration, as well as for screening modulators of these processes.
ARHGEF40 encodes a guanine nucleotide exchange factor (GEF) that specifically activates RhoA by catalyzing the exchange of GDP for GTP, yielding active RhoA-GTP. This activation is a critical node in the RhoA/ROCK signaling cascade, which governs actin cytoskeleton remodeling, cell migration, and proliferation. In these cells, ARHGEF40 functions downstream of TCR/CD28 co-stimulation and integrin-mediated adhesion, and can also be regulated by upstream receptor tyrosine kinases. Upon activation, RhoA-GTP engages downstream effectors including ROCK1/2, which phosphorylate LIMK, leading to cofilin inactivation, and myosin light chain 2 (MLC2), promoting actomyosin contraction. ARHGEF40 also interacts with ??-catenin and actin filaments, linking cytoskeletal dynamics to adhesive and transcriptional programs.
In the Jurkat T cell context, ARHGEF40-mediated RhoA activation is essential for proper immune synapse formation, sustained TCR signaling, and directed cell migration. Disruption of ARHGEF40 is predicted to impair RhoA-GTP loading, thereby attenuating ROCK-dependent cytoskeletal rearrangements required for T cell effector functions. This knockout model thus provides a powerful tool to dissect how TCR stimulation and integrin adhesion converge on RhoA to regulate immune cell polarity, migration, and cytokine production. Given the role of RhoA signaling in cancer metastasis and neurodevelopmental disorders, the ARHGEF40 knockout Jurkat cells also serve as a relevant platform for studying pathological cytoskeletal dysregulation.
Researchers can utilize these polyclonal knockout cells in a variety of advanced applications, including detailed analysis of TCR signaling kinetics, high-content imaging of immune synapse architecture, and quantitative assessment of cell migration using transwell assays. Representative experimental readouts include Western blotting for phospho-cofilin and phospho-MLC2, G-LISA to measure RhoA activation levels, F-actin staining with phalloidin, flow cytometric evaluation of activation markers CD69 and CD25, and co-immunoprecipitation to assess ARHGEF40?C??-catenin interactions. The polyclonal format is particularly suited for pooled screening assays, such as identifying chemical modulators of the RhoA/ROCK pathway, because it minimizes clonal artifacts and better represents the heterogeneous responses of a T cell population. For additional details or to request a quote, please contact Ascent Research.
