DOCK7 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HEK293T cells, designed for loss-of-function studies of the DOCK7 gene. This polyclonal population provides a genetically heterogeneous mixture of DOCK7-disrupted cells, enabling robust functional analysis without clonal isolation.
The host HEK293T cell line is a widely used human embryonic kidney epithelial model with high transfection efficiency and stable expression of the SV40 large T antigen, allowing episomal replication of plasmids containing the SV40 origin of replication. This background is ideal for transient and stable expression studies, viral packaging, and biochemical analyses.
DOCK7 encodes a guanine nucleotide exchange factor (GEF) that specifically activates the small GTPases Rac1 and Cdc42, serving as a critical link between upstream receptor tyrosine kinases (such as EGFR, ErbB2, and EphA2) and cytoskeletal reorganization. Upon stimulation, DOCK7-mediated GTP loading of Rac1/Cdc42 triggers downstream effectors including PAK1, JNK, and p38 MAPK, while also influencing the TSC1?CTSC2 complex to regulate mTORC1 signaling. Through these pathways, DOCK7 controls actin polymerization via LIMK and cofilin, as well as Arp2/3-mediated branching, driving cell migration, proliferation, and neurite outgrowth.
In the HEK293T background, loss of DOCK7 disrupts Rac1/Cdc42-dependent cytoskeletal dynamics and mTORC1 regulation, providing a clean system to dissect signaling crosstalk without confounding developmental factors. The high transfection efficiency facilitates complementation experiments, co-expression of pathway components, and rapid readouts of GTPase activity and actin reorganization. Because HEK293T cells endogenously express several upstream regulators (e.g., EGFR, ErbB2) and downstream effectors, the DOCK7 knockout allows for straightforward interrogation of receptor-to-mTORC1 and receptor-to-actin signaling cascades.
Researchers can employ these DOCK7 knockout polyclonal HEK293T cells in a variety of functional assays, including GTPase activation assays to measure Rac1/Cdc42 activity, cell migration and invasion assays, and phalloidin staining to visualize actin cytoskeletal changes. Western blot analysis of phospho-PAK1 and phospho-JNK provides a direct readout of signaling output, while co-immunoprecipitation of TSC2 assesses mTORC1 pathway engagement. The model is well suited for studying the molecular mechanisms underlying early infantile epileptic encephalopathy 23, skin hypopigmentation, and the role of DOCK7 in cancer progression. For additional details, please contact Ascent Research.
