The HS3ST1 Knockout KYSE-30 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the KYSE-30 human esophageal squamous cell carcinoma line. These cells harbor targeted disruption of the HS3ST1 gene, providing a robust loss-of-function model to investigate the biological roles of heparan sulfate 3-O-sulfation. The polyclonal format avoids single-cell cloning biases and preserves a heterogeneous pool of edited cells, making it suitable for a wide range of functional assays.
The parental KYSE-30 cell line was established from a poorly differentiated invasive esophageal squamous cell carcinoma resected from a 64-year-old Japanese male. KYSE-30 cells carry a TP53 mutation and are tumorigenic in immunocompromised mice, exhibiting characteristics of aggressive epithelial cancers. This line is extensively used in esophageal cancer research to study mechanisms of tumor progression, drug resistance, and interactions within the tumor microenvironment.
HS3ST1 encodes heparan sulfate glucosamine 3-O-sulfotransferase 1, an enzyme that catalyzes the 3-O-sulfation of glucosamine residues in heparan sulfate proteoglycans (HSPGs). This modification generates high-affinity binding sites for antithrombin, endowing HSPGs with anticoagulant properties, and for growth factors FGF2 and HGF, which are essential for FGF2?FGFR1 and HGF?MET signaling. HS3ST1 activity depends on the sulfate donor PAPS and is regulated by transcription factors ATF2 and NFKB1, as well as retinoic acid signaling. It functions in concert with other sulfotransferases, including HS2ST1 and HS3ST2, to produce defined sulfation patterns on core proteins such as syndecan-1 (SDC1) and glypican-1 (GPC1).
In the KYSE-30 esophageal carcinoma context, HS3ST1 knockout eliminates 3-O-sulfated heparan sulfate, thereby abrogating antithrombin binding and sequestering of FGF2 and HGF. Consequently, FGF2?induced phosphorylation of ERK and HGF?induced AKT activation are predicted to be diminished, leading to impaired proliferative and survival signals. Additionally, HSPG-mediated cell adhesion and migration are likely compromised. These alterations are expected to attenuate tumor growth, invasive capacity, and microenvironmental crosstalk, highlighting the knockout??s utility for dissecting the pro-tumorigenic functions of 3-O-sulfated heparan sulfate.
This polyclonal knockout cell product is engineered for diverse functional genomics applications. Researchers can validate HS3ST1 disruption via Western blotting, RT?qPCR, and immunofluorescence for 3-O-sulfated HS. Functional assays include antithrombin binding analysis to measure anticoagulant changes, FGF2?induced phospho?ERK ELISA to assess signaling output, and Boyden chamber-based migration/invasion assays to evaluate metastatic behavior. Transcriptomic profiling by RNA?seq enables global pathway discovery. These tools support investigations into heparan sulfate biosynthesis, growth factor signaling, anticoagulation biology, and esophageal squamous cell carcinoma therapeutics. For further information, please contact Ascent Research.
