The HS3ST1 Knockout SK-HEP-1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human liver adenocarcinoma cell line SK-HEP-1, featuring targeted disruption of the HS3ST1 gene. This heterogeneous pool of edited cells retains the genetic diversity of the parental line while eliminating HS3ST1 function, providing a loss-of-function model for studying 3-O-sulfation-dependent processes without single-cell cloning.
SK-HEP-1, originally isolated from ascites fluid of a liver adenocarcinoma patient, is a well-characterized endothelial-like cell line used as a model for liver sinusoidal endothelial cells and metastatic carcinoma. These cells display endothelial markers and tube-formation capacity, making them suitable for investigating hepatic cancer biology, tumor?Cendothelium interactions, and the role of glycosaminoglycans in the liver microenvironment. HS3ST1 knockout in this context enables dissection of specific heparan sulfate modifications in endothelial-like behavior.
HS3ST1 catalyzes 3-O-sulfation of glucosamine residues in heparan sulfate, generating the antithrombin III-binding motif. This modification is regulated by FGF2, TGF-beta, Wnt ligands, and BMP4, and operates downstream of the EXT1/EXT2 copolymerase complex, in coordination with NDST1, NDST2, and HS2ST. The resulting 3-O-sulfated domains activate antithrombin III for anticoagulation, modulate FGF2-FGFR1 signaling, and bind viral glycoproteins, impacting HSV-1 and HIV entry.
In SK-HEP-1 cells, HS3ST1 knockout ablates 3-O-sulfation, disrupting antithrombin III-binding and altering coagulation-related functions. This model is valuable for studying tumor-driven coagulation crosstalk, as cancers often exploit heparan sulfate for a prothrombotic niche. The endothelial-like properties facilitate analysis of altered FGF signaling, angiogenesis-related processes, and blood component interactions. The polyclonal population mirrors tumor heterogeneity, enhancing physiological relevance.
Research applications include anticoagulation mechanisms, cancer biology, and viral infection studies. Representative assays encompass western blotting for protein loss, RT-qPCR, heparan sulfate disaccharide analysis via HPLC-MS, antithrombin III binding and thrombin inhibition assays, HSV-1 entry assays, and FGF2 signaling assays. These cells support drug discovery targeting heparan sulfate?Cprotein interactions and glycosaminoglycan biosynthesis research. For further information, please contact Ascent Research.
