The GPR107 Knockout SK-HEP-1 Polyclonal Cells product provides a heterogeneous population of human hepatic adenocarcinoma cells engineered via CRISPR/Cas9-mediated disruption of the GPR107 gene. This polyclonal knockout model retains the genetic background of the SK-HEP-1 host cell line while introducing targeted loss-of-function mutations across the cell population, enabling functional studies without clonal selection artifacts. The polyclonal format reflects the spectrum of editing outcomes generated by CRISPR/Cas9, offering a robust and reproducible tool for investigating GPR107-dependent phenotypes in a liver cancer context. Researchers can utilize this model to dissect the role of GPR107 in intracellular trafficking, autophagy, and apoptotic regulation within a therapeutically relevant cellular system.
SK-HEP-1 is a well-characterized human cell line originally isolated from the ascites of a patient with liver adenocarcinoma. It displays an endothelial-like phenotype and is widely employed as a model for hepatic sinusoidal endothelium and hepatocellular carcinoma. The line’s dual epithelial and endothelial features make it particularly valuable for studying tumor microenvironment interactions, metastatic dissemination, and angiogenic signaling. In the context of GPR107 knockout, the SK-HEP-1 background allows exploration of how perturbations in endolysosomal trafficking influence liver cancer cell homeostasis, survival, and response to therapeutic stress.
GPR107 is an orphan G protein-coupled receptor localized predominantly to the trans-Golgi network and early endosomes, where it acts as a molecular scaffold for vesicular transport machinery. Mechanistically, it interacts with clathrin adaptor protein complexes AP-1 and AP-4 and the SNARE protein Vti1b to regulate cargo sorting and autophagic flux. Upstream signals such as nutrient deprivation and cellular stress activate GPR107-dependent pathways, which converge on downstream effectors including LC3, p62/SQSTM1, Caspase-3, and mTORC1. Disruption of GPR107 impairs autophagosome maturation and lysosomal degradation, leading to accumulation of LC3-II and p62, ultimately sensitizing cells to apoptotic cell death. The receptor thus represents a critical node connecting endosomal trafficking with autophagy and cell fate decisions.
In hepatic adenocarcinoma cells, GPR107 loss-of-function provides an experimentally tractable system to dissect how defects in intracellular trafficking and autophagy contribute to liver cancer pathogenesis. The SK-HEP-1 polyclonal knockout model enables investigation of GPR107??s role in sustaining cancer cell viability under metabolic stress and its potential as a therapeutic vulnerability. Because the polyclonal population harbors diverse editing events, it can reveal the continuum of phenotypic consequences associated with graded GPR107 inactivation, offering insights beyond those achievable with clonal knockouts. This model is particularly relevant for studying hepatocellular carcinoma, and may also inform research on neurodegenerative conditions in which autophagy dysfunction is implicated.
Typical applications include autophagic flux measurements using bafilomycin A1 and Western blotting for LC3 and p62 to monitor autophagosome turnover, apoptosis analysis by Annexin V/PI staining, and immunofluorescence localization of GPR107 and its interacting partners. Co-immunoprecipitation with AP-1 or Vti1b can probe trafficking complex integrity, while lysosomal activity and cell viability assays assess functional consequences of GPR107 disruption. These tools support primary research into autophagic cell death pathways, as well as drug screening for small molecules that modulate GPCR activity or restore lysosomal function. For further information or to discuss custom gene editing projects, please contact Ascent Research.
