This product consists of a CRISPR/Cas9-edited polyclonal knockout cell population of SK-OV-3 human ovarian adenocarcinoma cells, engineered to disrupt the HS3ST1 gene. The polyclonal format provides a heterogeneous pool of gene-edited cells, each carrying a genomic disruption of HS3ST1 introduced via non-homologous end joining, enabling loss-of-function studies without single-cell cloning. HS3ST1 encodes heparan sulfate 3-O-sulfotransferase 1, which catalyzes the transfer of sulfate groups to the 3-O position of glucosamine residues within heparan sulfate chains, a critical modification for generating binding motifs for antithrombin and viral glycoproteins. This knockout model serves as a robust tool for dissecting the functional roles of 3-O-sulfated heparan sulfate in cancer biology, viral entry, and anticoagulant signaling.
SK-OV-3 cells are an epithelial-like human ovarian adenocarcinoma cell line originally isolated from the ascites of a 64-year-old female. This line harbors mutations in TP53 and PIK3CA, making it a widely employed model for investigating ovarian cancer progression, metastasis, and drug resistance mechanisms. The SK-OV-3 background is particularly suited for examining heparan sulfate-dependent signaling due to its expression of proteoglycan core proteins such as syndecans and glypicans, which anchor heparan sulfate chains at the cell surface. In the context of ovarian cancer, HS3ST1-mediated 3-O-sulfation has been implicated in modulating growth factor responses and interactions with the tumor microenvironment, providing a relevant platform for functional interrogation.
The molecular network surrounding HS3ST1 involves its action on heparan sulfate precursor chains, which are synthesized by the copolymerase complex EXT1/EXT2 and sequentially modified by NDST1, HS2ST, and HS6ST. HS3ST1 is transcriptionally regulated by SOX2 and activated by the AP-1 complex, integrating its expression with broader oncogenic programs. The 3-O-sulfated motifs produced by HS3ST1 serve as high-affinity binding sites for antithrombin (SERPINC1), potentiating anticoagulant activity, and for herpes simplex virus glycoprotein D (gD), facilitating viral entry. In addition, these motifs modulate the binding of fibroblast growth factor 2 (FGF2) to its receptors, thereby influencing downstream ERK and AKT signaling cascades. Interacting proteins such as syndecans and glypicans present these modified heparan sulfate chains to the extracellular milieu, bridging extracellular cues to intracellular signal transduction.
Knockout of HS3ST1 in SK-OV-3 cells eliminates 3-O-sulfation of heparan sulfate, thereby disrupting critical receptor-ligand interactions that depend on this modification. This loss-of-function model impairs antithrombin binding, reducing the anticoagulant potential of the cell surface, and abrogates gD-mediated HSV-1 attachment, rendering cells resistant to viral entry. Concurrently, FGF2/ERK/AKT signaling is attenuated, which may affect cell proliferation, migration, and invasion??phenotypes central to ovarian cancer malignancy. By engineering HS3ST1 disruption in this TP53/PIK3CA-mutant background, researchers can specifically link 3-O-sulfate-dependent mechanisms to ovarian cancer pathophysiology without confounding genetic variables, enabling precise dissection of heparan sulfate??s role in oncogenic signaling and metastasis.
This polyclonal knockout cell population is suited for a diverse array of experimental applications. Researchers can employ HPLC-MS analysis to profile heparan sulfate disaccharide composition and confirm the loss of 3-O-sulfation, or utilize antithrombin affinity chromatography to quantify binding capacity. For signaling studies, FGF2-mediated phospho-ERK western blotting and downstream AKT pathway readouts are widely applicable. Functional assays include cell proliferation and wound healing migration tests to assess invasive behavior, as well as HSV-1 entry assays to evaluate viral susceptibility. Flow cytometry with anti-3-O-sulfated HS antibodies enables direct surface profiling, while qRT-PCR confirms HS3ST1 transcript ablation. This product is ideal for screening small-molecule HS3ST1 inhibitors, investigating heparan sulfate structure-function relationships in ovarian cancer, and studying viral entry mechanisms. For further technical details, please contact Ascent Research.
