The B3GALT6 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the HEK293T human embryonic kidney cell line, providing a loss-of-function model for investigating B3GALT6 in glycosaminoglycan (GAG) biosynthesis. This genetically disrupted model offers broad utility for studying proteoglycan biology without requiring single-cell cloning, maintaining robust target gene disruption across the cell pool.
HEK293T cells are widely used in biomedical research for their high transfectability, efficient recombinant protein expression, and lentiviral production capability. Originating from human embryonic kidney epithelium, they stably express SV40 large T antigen, facilitating episomal plasmid replication and high protein yields. This makes them a suitable model for proteoglycan synthesis and post-translational modification studies.
B3GALT6 encodes ??-1,3-galactosyltransferase 6, which transfers galactose to xylose in the proteoglycan tetrasaccharide linker, a crucial step in GAG chain initiation. It operates within a biosynthetic network alongside B4GALT7, B3GAT3, and the donor substrate UDP-galactose, downstream of xylosyltransferases XYLT1 and XYLT2 and upstream of phosphorylation and chain extension enzymes FAM20B and EXTL3. Disruption abolishes galactosylation, impairing heparan sulfate and chondroitin sulfate assembly, and disrupting growth factor signaling and cell-matrix interactions.
In HEK293T, B3GALT6 knockout allows detailed analysis of defective GAG linker biosynthesis. The cells endogenously express proteoglycan core proteins and GAG machinery components, enabling assessment of GAG composition, cell surface heparan sulfate, and effects on adhesion and receptor binding. This model is relevant to Ehlers-Danlos syndrome spondylodysplastic type, where B3GALT6 mutations cause linker defects leading to skeletal and skin abnormalities. Ablation recapitulates cellular disease phenotypes, aiding mechanistic studies and drug screening.
Applications include RT-qPCR for knockout confirmation, Western blot for glycosylation analysis, and dimethylmethylene blue (DMMB) assays for GAG quantification. Flow cytometry with heparan sulfate probes quantifies cell surface GAGs. The knockout also facilitates cancer glycosylation studies, where altered proteoglycan metabolism affects tumor progression and microenvironment signaling. It enables structure-function analyses of proteoglycan biosynthesis. For further details, contact Ascent Research.
