The HS3ST1 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for the study of heparan sulfate 3-O-sulfation in a human colorectal carcinoma background. This product provides a heterogeneous pool of HCT 116 cells in which the HS3ST1 gene has been disrupted, enabling loss-of-function analysis without clonal isolation. The polyclonal format preserves population-level diversity and serves as a robust tool for investigating the collective impact of HS3ST1 ablation on cellular phenotypes and signaling networks relevant to cancer biology and beyond.
HCT 116 is a widely used epithelial colorectal carcinoma cell line derived from an adult male patient. It exhibits an adherent growth pattern and is characterized by microsatellite instability, a KRAS G13D mutation, and a PIK3CA activating mutation. These genetic features render it a valuable model for colorectal cancer research, particularly for studying signaling pathways driven by oncogenic RAS and PI3K. The cell line’s origin from the colon and its well-characterized genomic landscape make it suitable for dissecting tumor-specific dependencies on heparan sulfate modifications.
HS3ST1 encodes heparan sulfate 3-O-sulfotransferase 1, which catalyzes the 3-O-sulfation of glucosamine residues within heparan sulfate chains. This modification is critical for generating high-affinity binding sites for antithrombin III and growth factors such as FGF2 and VEGF. HS3ST1 functions downstream of upstream regulators including TNF-alpha, TGF-beta, and EGF, which signal through NF-kB to modulate its expression. The 3-O-sulfated heparan sulfate chains interact with antithrombin III to accelerate coagulation inhibition, and with FGF2 and VEGF to promote FGFR- and VEGFR-mediated activation of ERK1/2 and AKT signaling. The biosynthetic pathway also involves EXT1, EXT2, and NDST1, which cooperate to build the heparan sulfate backbone prior to HS3ST1 action.
In the HCT 116 background, HS3ST1 knockout eliminates 3-O-sulfated domains, disrupting antithrombin binding and altering growth factor signaling balance. Given the host cell’s oncogenic KRAS and PIK3CA mutations, loss of HS3ST1 may attenuate FGF2- and VEGF-driven proliferation, migration, and survival signals, potentially affecting tumor progression and metastasis. This model also provides a platform to examine how altered cell surface heparan sulfate influences viral entry mechanisms, such as herpes simplex virus attachment, and coagulation-related interactions, linking colorectal cancer biology to thrombotic disorders.
Researchers can employ this knockout pool in a wide range of assays to explore heparan sulfate structure-function relationships. Western blotting and RT-qPCR confirm HS3ST1 loss, while mass spectrometry-based disaccharide analysis reveals changes in sulfation patterns. Functional studies may include FGF2 and antithrombin binding assays, phospho-ERK/AKT western blots to assess signaling output, and cell proliferation, migration, and invasion assays to evaluate phenotypic consequences. Flow cytometry with the 10E4 antibody detects cell surface heparan sulfate epitopes, and coagulation assays probe antithrombin-mediated activity. This model supports research in colorectal cancer, viral infections, and vascular biology. For further technical details and custom requirements, please contact Ascent Research.
