The HERC1 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human liver adenocarcinoma cell line SK-HEP-1. This product provides a heterogeneous pool of cells carrying targeted disruptions in the HERC1 gene, enabling loss-of-function studies without clonal selection. The polyclonal format preserves genetic diversity and reduces clonal artifacts, making it suitable for pooled screens and robust functional analyses. Researchers can interrogate the consequences of HERC1 ablation on ubiquitin-mediated signaling and cellular proliferation in a liver cancer context.
The parental SK-HEP-1 cell line was originally isolated from the ascites of a patient with liver adenocarcinoma. It serves as a well-established model for hepatic tumorigenesis and metastasis, exhibiting both epithelial and endothelial characteristics. SK-HEP-1 cells are widely employed to study hepatocellular carcinoma biology, metastatic progression, and drug responses. Their tumorigenic properties and liver origin make them an appropriate host for examining the role of ubiquitin ligases in hepatocarcinogenesis.
HERC1 encodes a large E3 ubiquitin ligase that catalyzes the covalent attachment of ubiquitin to substrate proteins, targeting them for degradation or modulating their activity. Biochemically, HERC1 interacts with and ubiquitinates TSC2, clathrin heavy chain, ARF1, and Rab GTPases, thereby regulating mTORC1 signaling, endosomal trafficking, and autophagy. It functions downstream of cellular stress signals, DNA damage, and nutrient deprivation, and is a key integrator of growth factor and metabolic cues. Through its interaction with mTORC1 components and the 26S proteasome, HERC1 controls protein homeostasis and cell cycle progression.
In the SK-HEP-1 host cell background, knockout of HERC1 is predicted to impair ubiquitination of TSC2, releasing inhibition of Rheb and resulting in constitutive mTORC1 hyperactivation. This dysregulation may drive abnormal cell proliferation, altered autophagy, and enhanced tumorigenic potential, mirroring aspects of hepatocellular carcinoma pathology. Given HERC1’s emerging role as a tumor suppressor in liver and colorectal cancers, this polyclonal knockout model enables dissection of mTOR-dependent and -independent mechanisms underlying its growth-regulatory functions.
Key applications include investigating HERC1’s tumor suppressor activity, characterizing mTOR pathway dysregulation in liver cancer, and conducting synthetic lethality screens to identify genes whose inactivation enhances HERC1-null cell death. The model is compatible with evaluation of drug sensitivity, particularly to mTOR inhibitors and ubiquitin pathway modulators. Typical assays include Western blotting for HERC1 and phosphorylated S6K, RT-qPCR for mTOR target genes, proliferation and colony formation assays, co-immunoprecipitation to assess TSC2 ubiquitination, immunofluorescence for endosomal markers, and drug sensitivity profiling. For further details or customization options, please contact Ascent Research.
