DPYSL2 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 hepatocellular carcinoma line. These cells, generated through CRISPR/Cas9-mediated gene disruption, provide a loss-of-function model for analyzing DPYSL2-dependent pathways. The polyclonal nature delivers a heterogeneous knockout population suitable for functional genomic studies. This knockout resource is designed for comprehensive functional investigations of DPYSL2 in a hepatocellular carcinoma context, aiding elucidation of its roles in cell migration, invasion, and cytoskeletal dynamics.
The Huh-7 cell line originates from a well-differentiated hepatocellular carcinoma of a Japanese male and retains hepatocyte-specific traits, including permissiveness to hepatitis C virus replication. It is a commonly used model for liver cancer biology, exhibiting relevant signaling pathways and morphologies essential for investigating migration, invasion, and cytoskeletal dynamics. It has been extensively utilized in liver cancer research to study tumor progression mechanisms.
DPYSL2 encodes CRMP2, a microtubule-associated protein mediating cytoskeletal reorganization. CRMP2 functions downstream of Semaphorin-3A/Neuropilin-1/PlexinA signaling and is phosphorylated by Cdk5 and GSK3??, thereby regulating microtubule assembly and actin polymerization. It interacts with tubulin, actin, kinesin-1, and focal adhesion components like paxillin and FAK, linking extracellular cues to cell adhesion and migration. Additionally, CRMP2 modulates the RhoA/ROCK pathway, influencing cofilin activity and actin turnover.
In Huh-7 hepatocellular carcinoma cells, DPYSL2 knockout disrupts coordinated cytoskeletal dynamics, impairing cell motility and invasion??key traits of metastatic cancer. This model allows dissection of CRMP2??s role in liver cancer-specific signaling, including responses to Sema3A and growth factor stimulation, and its effect on downstream effectors such as LIMK and cofilin. Consequently, these cells provide a platform to probe the contribution of DPYSL2 to hepatocarcinogenesis.
These polyclonal knockout cells are ideal for transwell migration and Matrigel invasion assays, immunofluorescence staining of tubulin and F-actin, and RhoA activity measurements. They support proliferation assays (e.g., MTT), western blotting and RT-qPCR for DPYSL2 validation, and co-immunoprecipitation for protein interaction studies. Researchers can employ these cells to assess the impact of DPYSL2 ablation on tumor cell behavior and signaling transduction. Furthermore, they serve as a valuable tool for drug target validation and screening of compounds targeting CRMP2-related pathways. For further information, contact Ascent Research.
