The ARPC5L Knockout Jurkat Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T lymphocyte line, engineered to disrupt the ARPC5L gene. This pooled knockout model provides a robust tool for studying loss-of-function effects of ARPC5L, a key subunit of the Arp2/3 complex, without clonal isolation. The polyclonal format preserves genetic heterogeneity while ensuring targeted gene disruption, making it suitable for population-level analyses of actin cytoskeleton dynamics and T-cell functions. The cells are supplied as a polyclonal knockout population, ready for downstream functional assays, and are validated for ARPC5L protein loss by Western blotting.
Jurkat cells are an extensively characterized human T lymphocyte line originally derived from the peripheral blood of a male patient with T-cell acute lymphoblastic leukemia (T-ALL). They serve as a critical model system for investigating T-cell receptor (TCR) signaling, activation, apoptosis, and leukemia biology. These suspension cells exhibit constitutive T-lymphoblastoid properties and recapitulate key aspects of early T-cell signaling events, including calcium flux, kinase cascades, and transcriptional responses. Their well-defined genetic background and ease of transfection or electroporation make them a preferred host for gene editing studies, particularly for exploring actin-mediated processes during immune cell activation and migration.
ARPC5L encodes the ARPC5-like subunit of the Arp2/3 complex, which promotes actin filament nucleation and branching, essential for cytoskeletal remodeling. This complex is activated by nucleation-promoting factors such as WAVE2, functioning downstream of Rac1 and Cdc42 Rho GTPases, and integrates signals from TCR activation and Src family kinases. ARPC5L interacts directly with other core subunits, including ARPC1A, ARPC2, ARPC3, ARPC4, ACTR2, and ACTR3, and associates with regulatory proteins like cortactin and vinculin. Downstream, Arp2/3-mediated actin polymerization drives lamellipodia formation, cell migration, endocytosis, integrin clustering, and the actin reorganization required for immune synapse formation. Mechanistically, ARPC5L loss disrupts this nucleation, impairing actin network assembly at the leading edge and compromising signal transduction to MRTF-A and SRF transcriptional programs.
In the Jurkat T-cell context, ARPC5L knockout profoundly affects actin-dependent processes critical for lymphocyte function. The disruption of Arp2/3-mediated actin nucleation impairs lamellipodia extension and cell polarization, leading to reduced chemotactic migration. Moreover, TCR-induced actin remodeling at the immune synapse is compromised, altering the spatial organization of signaling molecules and dampening downstream phospho-signaling events, such as ZAP70 and ERK phosphorylation. This model therefore enables dissection of the specific requirement for ARPC5L in linking TCR-mediated Rac1/WAVE2 activation to cytoskeletal effector responses, with relevance to understanding actin-related immunodeficiencies, WASP signaling defects, and T-ALL pathobiology.
Researchers utilize these ARPC5L knockout Jurkat polyclonal cells for a wide array of assays, including immunofluorescence staining with phalloidin to visualize F-actin structures, live-cell imaging of actin dynamics, flow cytometry-based adhesion and migration assays, and T-cell activation assays measuring CD69 upregulation. Western blotting confirms ARPC5L absence, and co-immunoprecipitation reveals alterations in Arp2/3 complex composition. Applications extend to chemotaxis assays, phospho-signaling analyses, and drug screening for actin-targeting compounds. This model supports studies of cancer metastasis, developmental disorders of the actin cytoskeleton, and immune dysregulation, providing a physiologically relevant platform for mechanistic and translational research. For additional product details or technical support, please contact Ascent Research.
