The DOCK4 knockout A-549 polyclonal cell product consists of a population of A-549 lung adenocarcinoma epithelial cells with CRISPR/Cas9-mediated disruption of the DOCK4 gene. This polyclonal knockout cell population is designed for loss-of-function studies of DOCK4, a guanine nucleotide exchange factor (GEF) critical for Rac1 and Rap1 activation. The knockout model enables researchers to dissect the role of DOCK4 in cell migration, adhesion, and cytoskeletal dynamics.
The A-549 cell line is a well-characterized model of human lung adenocarcinoma, originally isolated from a 58-year-old Caucasian male. These cells exhibit features of alveolar type II pneumocytes and are widely used to investigate lung cancer biology, including proliferation, migration, and metastasis. The epithelial origin and tumorigenic properties of A-549 cells provide a physiologically relevant context for studying DOCK4-dependent pathways in non-small cell lung cancer.
DOCK4 functions as a GEF that activates the small GTPases Rac1 and Rap1 by catalyzing GDP-to-GTP exchange. Upon integrin engagement or growth factor stimulation via receptors such as EGFR and PDGFR, DOCK4 is recruited to the plasma membrane through interaction with ELMO2. Activated Rac1 triggers downstream effectors including PAK, the WAVE complex, Arp2/3, and LIMK/cofilin, leading to actin polymerization, lamellipodia formation, and focal adhesion turnover. DOCK4 also interacts with NCK2 and Integrin ??1 and is regulated by PI3K-dependent signals. These molecular interactions place DOCK4 at the nexus of integrin and receptor tyrosine kinase signaling cascades that coordinate cell motility.
In A-549 lung adenocarcinoma cells, DOCK4-mediated Rac1 activation promotes migratory and invasive phenotypes characteristic of metastatic cancer. Disruption of DOCK4 expression in this model allows researchers to interrogate how loss of Rac1/Rap1 GEF activity affects matrix degradation, cell-substrate adhesion strength, and directional migration. This polyclonal knockout population is particularly suitable for examining the role of DOCK4 in epithelial-mesenchymal transition (EMT) processes and for validating DOCK4 as a potential therapeutic target in lung adenocarcinoma.
Typical applications include scratch wound healing assays and Transwell Matrigel invasion assays to quantify migration and invasion defects. Rac1-GTP pull-down activation assays can assess GTPase activity, while immunofluorescence staining for F-actin, paxillin, and vinculin reveals changes in cytoskeletal organization and focal adhesion dynamics. Western blotting for DOCK4, ELMO2, and downstream phospho-targets confirms knockout efficiency and pathway alterations. This product is also valuable for drug target validation studies aimed at developing inhibitors of the DOCK4-ELMO2-Rac1 signaling axis. For further technical information or to discuss custom orders, please contact Ascent Research.
