DOCK5 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human hepatocellular carcinoma Huh-7 cell line. This product provides a mixed population of cells carrying disruptions in the DOCK5 gene, enabling loss-of-function studies without clonal selection. The CRISPR/Cas9-mediated gene disruption targets the DOCK5 locus, creating a heterogeneous knockout model that preserves the genetic diversity of the parental line while abrogating DOCK5 protein function.
Huh-7 is a well-characterized hepatic epithelial cell line originally established from a liver tumor of a 57-year-old Japanese male with hepatocellular carcinoma. These cells maintain key features of hepatocyte-like morphology and express markers relevant to liver cancer biology, including alpha-fetoprotein. Huh-7 cells are widely employed in the study of hepatocarcinogenesis, drug metabolism, and metastatic progression, making them an appropriate platform for investigating the role of DOCK5 in liver cancer cell motility and invasion.
DOCK5 encodes a guanine nucleotide exchange factor (GEF) that specifically activates Rac1 by catalyzing the exchange of GDP for GTP. In complex with the adapter proteins ELMO1 and ELMO2, DOCK5 promotes Rac1-GTP loading and subsequent activation of downstream effectors such as PAK1 and LIMK1, which regulate cofilin-mediated actin reorganization. This signaling is triggered by upstream cues from HGF/MET, integrin-mediated adhesion, and PI3K pathways, linking extracellular stimuli to cytoskeletal rearrangements. DOCK5-mediated Rac1 activation drives lamellipodia formation and focal adhesion turnover, critical processes for cell migration and invasion.
In hepatocellular carcinoma, elevated DOCK5 expression correlates with enhanced metastatic potential. Knockout of DOCK5 in Huh-7 cells abrogates Rac1-mediated actin polymerization and lamellipodia formation, leading to impaired cell migration and invasion. This polyclonal knockout model recapitulates the loss of DOCK5 function in a heterogeneous background, reflecting the clonal diversity present in tumors. Consequently, these cells are a valuable tool for dissecting the DOCK5-ELMO-Rac1 signaling axis in liver cancer and for evaluating the impact of pathway inhibition on metastatic behavior without the confounding effects of clonal selection.
Researchers can employ DOCK5 Knockout Huh-7 Polyclonal Cells in a range of functional assays, including Transwell migration and invasion assays, wound healing assays, and cell adhesion assays to assess the role of DOCK5 in metastatic processes. The cells are also suitable for Rac1-GTP pull-down experiments to quantify Rac1 activation levels and immunofluorescence staining for F-actin and paxillin to visualize actin cytoskeleton changes. These applications support studies aimed at elucidating molecular mechanisms of hepatocellular carcinoma metastasis and screening anti-metastatic therapeutic candidates. For further information or to discuss custom projects, please contact Ascent Research.
