The ITSN2 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population derived from the human hepatic adenocarcinoma SK-HEP-1 cell line, with targeted disruption of the ITSN2 gene. This loss-of-function model is generated using a polyclonal format, ensuring a heterogeneous knockout pool that reflects the complexity of genetic perturbation.
The SK-HEP-1 host cell line was originally established from the ascitic fluid of a male patient diagnosed with hepatic adenocarcinoma. As a widely utilized model, SK-HEP-1 cells display characteristics of mesenchymal origin yet are employed in hepatocellular carcinoma research, offering a platform to study liver cancer cell biology, metastasis, and therapeutic responses.
ITSN2 encodes a multidomain scaffold protein that orchestrates clathrin-mediated endocytosis, actin cytoskeleton remodeling, and receptor tyrosine kinase signal transduction. The protein interacts directly with dynamin, synaptojanin, Cbl, Eps15, epsin, and N-WASP, coupling endocytic vesicle formation to actin polymerization. ITSN2 operates downstream of epidermal growth factor (EGF) and other receptor tyrosine kinases, linking EGFR internalization to downstream MAPK pathway activation. Mechanistically, ITSN2 facilitates the recruitment of Cbl and Eps15 to activated EGFR, promoting receptor ubiquitination, endocytosis, and subsequent signaling through dynamin-dependent vesicle scission and actin filament dynamics.
In the SK-HEP-1 hepatic adenocarcinoma context, disruption of ITSN2 profoundly alters EGFR-mediated signaling and cellular behavior. The knockout model is characterized by impaired internalization of EGFR and attenuated phosphorylation of ERK, a key MAPK effector, resulting in diminished cell migration and proliferation. This phenotype underscores the critical role of ITSN2 in coupling endocytic trafficking to proliferative and migratory signals in liver cancer cells, making the polyclonal knockout population a relevant system for dissecting ITSN2-dependent oncogenic mechanisms.
This ITSN2 knockout polyclonal cell product is suited for a diverse array of functional studies, including investigation of endocytosis-dependent signaling in hepatocellular carcinoma, CRISPR-based functional genomics, and drug sensitivity profiling. Researchers can employ western blotting and RT-qPCR to confirm ITSN2 ablation, immunofluorescence and flow cytometry to assess EGFR trafficking, and wound healing or invasion assays to quantify migratory changes. Phospho-ERK analysis and EGFR internalization assays offer precise readouts of signaling perturbation, while proliferation and drug sensitivity assays facilitate therapeutic evaluation in the context of liver cancer. For further details and ordering information, please contact Ascent Research.
