The DOCK4 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the gene encoding dedicator of cytokinesis 4 (DOCK4) has been disrupted within the HEK293T host background. This product provides a heterogeneous pool of cells harboring targeted loss-of-function modifications in the DOCK4 locus, enabling the study of DOCK4-dependent signaling and cellular processes without the isolation of a monoclonal derivative. The polyclonal format preserves population-level genetic diversity while eliminating DOCK4 protein expression, making it suitable for experiments where clonal variability is to be minimized or where bulk assays are preferred.
HEK293T cells are a widely employed human embryonic kidney epithelial line that stably expresses the SV40 large T antigen, which facilitates episomal replication of plasmids containing the SV40 origin of replication and enhances recombinant protein production. Their high transfection efficiency, rapid growth, and robust expression of exogenous constructs have established HEK293T as a versatile model for biochemical, cell biological, and pharmacological investigations. The epithelial origin and adherent nature of these cells support studies of cell morphology, adhesion, and migration, and they retain key signaling modules relevant to oncogenic and developmental pathways.
DOCK4 functions as a guanine nucleotide exchange factor (GEF) that specifically activates the small GTPases Rac1 and Rap1, thereby orchestrating actin cytoskeleton reorganization, cell migration, and adhesion. It operates through a conserved DOCK-homology region 2 (DHR2) catalytic domain and forms a regulatory complex with the engulfment and cell motility proteins ELMO1 and ELMO2. Upstream inputs include growth factor stimulation (e.g., EGF), integrin engagement, GPCR ligands, and the small GTPase RhoG. Upon activation, DOCK4 promotes GTP loading of Rac1 and Rap1, leading to downstream engagement of PAK kinases, the WAVE regulatory complex, Arp2/3-mediated actin nucleation, and MAPK cascades. This signaling axis couples extracellular cues to dynamic changes in the cytoskeleton and focal adhesion turnover, positioning DOCK4 as a critical node in pathways regulating directional cell motility and tissue morphogenesis.
The HEK293T cell model provides an experimentally tractable system to interrogate DOCK4 function because these cells exhibit active Rac1 and Rap1 signaling and readily form lamellipodia and focal adhesions upon stimulation. Disruption of DOCK4 in this background creates a loss-of-function platform to dissect the molecular contributions of DOCK4 to GTPase activation, cytoskeletal remodeling, and migration independently of other DOCK family members. Due to the polyclonal nature of the knockout, the population can be directly compared to parental HEK293T cells in quantitative biochemical and imaging assays, revealing the net effect of DOCK4 ablation on signaling networks and cellular behavior.
This DOCK4 polyclonal knockout product is intended for a broad range of research applications, including cellular migration and invasion assays (e.g., wound healing and Transwell invasion), Rac1 and Rap1 activity measurements by G-LISA or pull-down, immunofluorescence staining of F-actin to assess cytoskeletal organization, co-immunoprecipitation of ELMO?CDOCK4 complexes, and RT-qPCR analysis of downstream transcriptional targets. The cells are also valuable for modeling GPCR-mediated Rac activation and investigating the role of DOCK4 in cancer cell dissemination and neurodevelopmental processes. For additional details or technical support, please contact Ascent Research.
