The HS3ST1 Knockout CAL-27 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population derived from the human oral squamous cell carcinoma line CAL-27, engineered to disrupt the HS3ST1 gene. This knockout model abolishes the expression of heparan sulfate 3-O-sulfotransferase 1, enabling investigation of 3-O-sulfated heparan sulfate motifs in epithelial cancer biology. The polyclonal nature preserves population-level heterogeneity while ensuring target gene disruption, making it suitable for bulk functional genomics and phenotypic assays.
The CAL-27 host cell line was established from a tongue squamous cell carcinoma of a 56-year-old male patient and exhibits adherent epithelial growth. CAL-27 is widely employed as a model for head and neck squamous cell carcinoma (HNSCC), reflecting key molecular alterations in oral carcinogenesis. Its use in knockout studies allows dissection of gene function in a clinically relevant cancer context, particularly those linked to extracellular matrix remodeling, growth factor signaling, and invasion.
HS3ST1 encodes the enzyme that catalyzes the transfer of sulfate to the 3-O position of glucosamine residues within heparan sulfate chains, a critical modification that creates high-affinity binding sites for antithrombin III, thus enhancing anticoagulant activity. This sulfation event also modulates fibroblast growth factor 2 (FGF2) signaling by altering interactions between heparan sulfate proteoglycans (HSPGs; e.g., syndecans, glypicans) and FGF receptors (FGFRs). Upstream regulators such as FGF2 and TGF-?? signaling pathways influence HS3ST1 expression, while downstream targets include antithrombin III?Cthrombin complexes and phospho-ERK cascades. The enzyme functions within a multiprotein biosynthetic complex containing EXT1/EXT2 copolymerases and can be influenced by growth factors like VEGF.
In the context of oral squamous cell carcinoma, HS3ST1-mediated heparan sulfate sulfation may contribute to tumor progression by fine-tuning growth factor gradients and cell?Cmatrix interactions. CAL-27 cells with HS3ST1 knockout provide a model to assess how loss of 3-O-sulfation impacts proliferative signaling, migration, and responses to therapeutic agents. Given the role of HSPGs in co-receptor functions, this knockout system can reveal dependencies on specific sulfation patterns that might be exploited therapeutically.
This polyclonal knockout cell population is suited for a range of applications, including HPLC-MS disaccharide analysis to profile heparan sulfate fine structure, antithrombin binding ELISAs to quantify functional interactions, phospho-ERK western blotting to measure FGF2-dependent signaling, and flow cytometry with anti-heparan sulfate antibodies to assess global sulfation changes. Viral entry studies using HSV-1 glycoprotein D binding assays can explore pathogen?Chost glycocalyx interactions, while Sanger sequencing confirms on-target gene disruption. For further information or assistance, please contact Ascent Research.
