The HPS3 Knockout SK-HEP-1 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human liver adenocarcinoma cell line, engineered for targeted disruption of the HPS3 gene. This polyclonal knockout pool offers a loss-of-function model for investigating HPS3-dependent pathways without the need for single-cell cloning, providing a heterogeneous mixture of edited alleles representative of a population-level knockout effect.
The parental SK-HEP-1 cell line originates from the ascitic fluid of a patient with liver adenocarcinoma and is extensively employed as an in vitro model for liver sinusoidal endothelial cells, despite its epithelial origin. This cell line retains characteristics relevant to hepatic endothelial biology, including the expression of certain endothelial markers, making it suitable for studies of liver-specific vesicular trafficking and organelle dynamics.
HPS3 encodes a subunit of the biogenesis of lysosome-related organelles complex-2 (BLOC-2), forming a functional module with HPS5 and HPS6 that mediates cargo transport from early endosomes to lysosome-related organelles, including melanosomes and platelet dense granules. The complex acts downstream of AP-3 and cooperates with Rab32 and Rab38 to deliver melanosomal proteins such as TYR and TYRP1, lysosomal enzymes, and dense granule components. HPS3 knockout abrogates this trafficking, disrupting organelle maturation.
In the SK-HEP-1 hepatic endothelial-like context, HPS3 knockout allows dissection of BLOC-2-dependent trafficking specific to this cell type. It also permits examination of how defective organelle biogenesis intersects with hepatocellular carcinoma biology, given the cell line??s dual epithelial?Cendothelial phenotype. The model is pertinent to studying platelet dense granule and melanosome defects underlying the bleeding diatheses and albinism of Hermansky-Pudlak syndrome type 3.
Typical applications include immunofluorescence for organelle markers, Western blotting of BLOC-2 components, electron microscopy, platelet dense granule ATP release assays, and melanosome maturation assays. Co-immunoprecipitation can assess complex integrity, and RT-qPCR verifies gene disruption. Thus, these cells are valuable for Hermansky-Pudlak syndrome research, organelle biogenesis studies, pigmentation disorder investigations, and platelet granule formation analyses. For further information or to discuss customized knockout solutions, please contact Ascent Research.
