The CD2AP Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the CD2AP gene in the SK-HEP-1 hepatocellular carcinoma background. Supplied as a non-clonal knockout pool, this model avoids the clonal selection biases inherent in single-cell-derived lines, providing a physiologically relevant loss-of-function system. CD2AP encodes an adaptor protein that scaffolds membrane receptors to the actin cytoskeleton and endocytic machinery; its ablation enables systematic analysis of receptor trafficking, cytoskeletal dynamics, and downstream signaling pathways.
SK-HEP-1 is a liver adenocarcinoma cell line derived from ascitic fluid and characterized by a unique hybrid endothelial-hepatocyte phenotype. It co-expresses epithelial and mesenchymal markers, making it an excellent model for studying epithelial-mesenchymal transition, migration, and adhesion. Its metastatic origin and capacity for rapid growth facilitate investigations into aggressive cancer cell biology and signal transduction. These features render SK-HEP-1 particularly suited to interrogate CD2AP functions in actin remodeling, endocytosis, and mTOR signaling within a cancer-relevant context.
CD2AP functions as a critical adaptor linking transmembrane proteins such as nephrin, CD2, and the EGF receptor to the actin polymerization machinery and endosomal trafficking regulators. Through direct interactions with cortactin, CAPZA1, Rab4, and Rab5, CD2AP orchestrates Arp2/3?Cmediated actin nucleation via N-WASP and modulates Rac1 activity. It also forms the nephrin?CCD2AP?Cpodocin slit diaphragm complex in podocytes, a module that can be reconstituted heterologously. Activation of mTORC1 is controlled through CD2AP-dependent endosomal sorting of growth factor receptors. Disruption of CD2AP breaks these connections, impairing actin dynamics, endocytosis, and signal integration.
Within the SK-HEP-1 background, CD2AP knockout impairs migration, invasion, and endocytic recycling, processes essential for hepatocellular carcinoma metastasis. Decoupling of integrin and EGFR signaling from the cytoskeleton disrupts adhesion and motility. This model facilitates dissection of pathways underlying focal segmental glomerulosclerosis, nephrotic syndrome, and Alzheimer??s disease. It is an effective tool for analyzing CD2AP?Ccortactin?CArp2/3, Rab endosome networks, and mTORC1 regulation in a hepatic tumor setting.
Typical investigations include assessment of protein expression by western blot and immunofluorescence, endocytosis kinetics using fluorescent cargo uptake, and migration/invasion via transwell assays. Co-immunoprecipitation probes altered protein interactions, while RT-qPCR and flow cytometry quantify transcriptional and proliferation changes. Although hepatic in origin, SK-HEP-1 cells accommodate podocyte-relevant co-expression studies, enabling kidney disease modeling. The polyclonal format ensures population-level data with minimal clonal artifacts. For ordering and technical inquiries, please reach out to Ascent Research.
