The DOCK10 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the DOCK10 gene has been disrupted. This product provides a loss-of-function model for investigating the roles of DOCK10, a guanine nucleotide exchange factor (GEF) specific for the small GTPase Cdc42. The polyclonal format preserves population heterogeneity while eliminating functional DOCK10 protein, enabling researchers to examine DOCK10-dependent phenotypes in a robust, cost-effective manner without the need for single-cell cloning or biallelic validation. This model is ideal for studying DOCK10-mediated signaling cascades and cellular processes in an experimentally tractable human cell background.
HEK293T cells are a widely used human embryonic kidney epithelial line immortalized by constitutive expression of the SV40 large T antigen, which facilitates high-yield protein production and efficient transfection. Their adherent, epithelial morphology and well-characterized signaling networks make them a versatile platform for dissecting cytoskeletal dynamics and GTPase regulation. Although not of immune origin, HEK293T cells express core machinery for Rho GTPase signaling and actin remodeling, providing a relevant context for studying core migratory and morphological pathways that are conserved across cell types. This cell line is routinely employed in studies of signal transduction, protein interaction, and cell migration assays.
DOCK10 encodes an atypical DOCK-family GEF that activates Cdc42 by catalyzing the exchange of GDP for GTP. Active, GTP-bound Cdc42 then transduces signals to downstream effectors including PAK1 and WASP, which drive actin polymerization and formation of lamellipodia and filopodia. DOCK10 is activated by upstream cues such as integrin signaling, chemokine receptors, and PI3K, and it operates within a broader Rho GTPase cycle that includes Rac, PAK, LIMK, and cofilin. DOCK10 forms functional complexes with ELMO1, ELMO2, and the small GTPase RHOG, which help localize and stabilize its activity at membrane ruffles. Through this Cdc42-PAK-WASP axis, DOCK10 fundamentally regulates actin cytoskeleton reorganization, cell spreading, and directed migration.
In HEK293T cells, disruption of DOCK10 impairs the normal activation of Cdc42, leading to attenuated phosphorylation of downstream PAK kinases and diminished actin remodeling. This loss-of-function context enables dissection of DOCK10-specific contributions to cell motility, adhesion dynamics, and cytoskeletal architecture without confounding effects from closely related GEFs. The model allows direct interrogation of DOCK10??s role in linking extracellular stimuli??such as integrin engagement or chemokine cues??to Cdc42-driven morphological responses. Researchers can compare parental and knockout populations in a range of quantitative migration and invasion assays to isolate DOCK10-dependent events.
The DOCK10 Knockout HEK293T Polyclonal Cells are well-suited for investigating Cdc42 signaling networks, Rho GTPase regulation, and actin dynamics. Typical applications include pull-down assays for GTP-Cdc42 to measure direct Cdc42 activation, Western blotting for phosphorylated PAK and other effectors, transwell migration and Matrigel invasion assays, and immunofluorescence microscopy to visualize F-actin structures and cell spreading. These tools support research into cancer metastasis, immune cell migration, and related disorders. For further details and ordering information, please contact Ascent Research.
