The DOCK4 Knockout HGC-27 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of HGC-27 human gastric carcinoma epithelial cells with a targeted disruption of the DOCK4 gene. This loss-of-function model is generated using CRISPR/Cas9 nuclease technology to introduce gene-inactivating mutations across the cell pool, without the need for single-cell cloning. The polyclonal format ensures representative knockout efficiency while maintaining the genetic heterogeneity of the parental line, making it ideal for population-based functional assays.
HGC-27 cells were derived from the lymph node metastasis of an undifferentiated gastric adenocarcinoma and serve as a well-characterized in vitro model of metastatic gastric cancer. They display epithelial morphology, robust proliferation, and migratory properties that closely mirror aggressive disease. This background provides a clinically relevant system to study the molecular determinants of gastric cancer progression and distant spread.
DOCK4 is an atypical guanine nucleotide exchange factor (GEF) for the small GTPase RAC1, functioning in a complex with ELMO1 or ELMO2. It catalyzes GDP/GTP exchange on RAC1, leading to downstream activation of PAK, JNK, and p38 MAPK, and ultimately orchestrating actin cytoskeleton remodeling through the LIMK-cofilin and Arp2/3 pathways. Upstream integrin and growth factor receptor signals converge on DOCK4 to regulate lamellipodia formation, cell adhesion, and directed migration. In gastric cancer, DOCK4 has been implicated as a tumor suppressor, and its loss correlates with enhanced invasive behavior.
Disruption of DOCK4 in HGC-27 cells allows direct investigation of its tumor-suppressive functions within a gastric cancer context. Knockout of DOCK4 is expected to alter RAC1?mediated actin dynamics, potentially increasing cell motility and invasive capacity. This model facilitates the dissection of DOCK4-dependent signaling networks and their interplay with pathways such as Wnt signaling, providing insights into mechanisms of metastasis. By comparing CRISPR-edited knockout cells to parental controls, researchers can define the role of DOCK4 in processes like epithelial-mesenchymal transition and matrix degradation.
Key applications include transwell migration and invasion assays, wound healing assays, and RAC1 activation measurements using G-LISA or pull-down approaches. Downstream signaling can be assessed by western blotting for phospho-PAK and phospho-cofilin, while immunofluorescence reveals changes in F-actin organization. Co-immunoprecipitation studies confirm DOCK4-ELMO complex integrity. The knockout cells are also suitable for RNA-seq transcriptome profiling and high-throughput screening of anti-metastatic compounds. Proliferation assays further augment functional characterization. For further details or technical inquiries, contact Ascent Research.
