The B3GAT3 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting B3GAT3 in human HEK293T cells. This loss-of-function model enables study of glycosaminoglycan biosynthesis by disrupting the glucuronosyltransferase I responsible for completing the tetrasaccharide linker of proteoglycans. The heterogeneous knockout pool, generated via CRISPR/Cas9-mediated gene disruption, minimizes clonal bias and is ideal for investigating collective effects of B3GAT3 deficiency on heparan sulfate and chondroitin sulfate chain assembly.
Derived from human embryonic kidney epithelial cells, HEK293T cells are immortalized with SV40 large T antigen, conferring high transfection efficiency and robust protein expression capability. Widely used for viral packaging and recombinant protein production, these cells provide a well-characterized epithelial background with endogenous glycosylation machinery suitable for studying proteoglycan metabolism. The knockout thus offers a tractable platform to examine post-translational modifications in a simplified system.
B3GAT3 (glucuronosyltransferase I) catalyzes glucuronic acid addition to the Gal-Gal-Xyl-Ser linker, acting downstream of B3GALT6 and upstream of EXT1/EXT2 and CHSY1/CHPF. It interacts with glycosyltransferases B4GALT7, B3GALT6, XYLT1, and XYLT2, and its substrate xylose is phosphorylated by FAM20B. Knockout of B3GAT3 abrogates polymerization of heparan sulfate onto syndecans and glypicans, as well as chondroitin sulfate assembly, thereby disrupting growth factor signaling, adhesion, and matrix interactions dependent on these glycosaminoglycans.
In HEK293T cells, the loss of functional glycosaminoglycans allows clean dissection of signaling pathways that require heparan sulfate co-receptors, such as FGF, Wnt, and Hedgehog cascades active in developing kidney epithelia. The model also aids viral entry studies, as many viruses exploit cell-surface heparan sulfate for attachment; B3GAT3 knockout abolishes this glycan, clarifying receptor requirements. This specific genetic ablation provides a definitive link between linker formation and downstream phenotypes.
Applications encompass functional studies of heparan sulfate-dependent growth factor responses via phospho-proteomic analysis, modeling congenital disorders like Larsen-like syndrome through cell migration assays, and evaluating viral tropism. Compatible methods include immunofluorescence for glycosaminoglycan epitopes, flow cytometry, HPLC disaccharide profiling, and Alcian blue staining for sulfated proteoglycans. For further details, please contact Ascent Research.
