ABR Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 human hepatocellular carcinoma cell line with disruption of the ABR gene. This polyclonal pool serves as a loss-of-function model to study ABR??s role in actin cytoskeleton regulation and cell motility. ABR encodes a GTPase-activating protein for RAC1 and CDC42, and its knockout is predicted to enhance RAC/CDC42 activity, making it valuable for cancer metastasis research.
Huh-7 is an epithelial cell line isolated from a liver tumor of a 57-year-old Japanese male, widely used in hepatitis C virus and hepatocellular carcinoma studies. These tumorigenic cells retain metabolic and detoxification functions, providing a robust platform to investigate liver cancer signaling pathways, drug responses, and metastatic mechanisms.
Mechanistically, ABR inactivates RAC1 and CDC42 by accelerating GTP hydrolysis. It is regulated by integrin signaling and growth factor receptors like EGFR, and it interacts with 14-3-3 proteins, BCR, and NCK1. Downstream, ABR modulates PAK1, WASP, and the Arp2/3 complex, controlling actin polymerization and cell adhesion. This positions ABR at a nexus between extracellular cues and cytoskeletal remodeling, influencing LIMK and cofilin activity.
In Huh-7 cells, ABR knockout is expected to increase cell migration and invasion, processes central to hepatic metastasis. This model enables examination of how ABR integrates signals to regulate hepatocellular carcinoma cell dynamics, providing a tool to dissect ABR??s impact on tumorigenic potential and cytoskeletal dysregulation.
Applications include Boyden chamber migration and invasion assays, phalloidin-based actin staining, and adhesion experiments. Mechanistic studies can employ western blotting for RAC1-GTP, CDC42-GTP, and phospho-PAK, along with Rho GTPase activation pull-downs. The model supports RNA-seq, xenograft tumor models, and drug screening for metastasis inhibitors. Contact Ascent Research for additional details.
