The B3GALT6 Knockout HT29 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout population with targeted gene disruption of B3GALT6 in the HT29 colorectal adenocarcinoma epithelial cell line. This heterogeneous loss-of-function model is ideal for functional studies of proteoglycan biosynthesis and glycosaminoglycan (GAG) chain initiation without requiring clonal selection.
HT29 cells, originally derived from a primary colorectal adenocarcinoma of a 44-year-old Caucasian female, are a standard intestinal epithelial model capable of enterocyte-like differentiation. They are widely used in colorectal cancer research for investigating barrier function, drug absorption, and tumor biology, providing a physiologically relevant background to examine B3GALT6-dependent glycosylation effects.
B3GALT6 encodes beta-1,3-galactosyltransferase II, which catalyzes the addition of the second galactose residue to the GAG linker tetrasaccharide, a critical step shared by heparan sulfate and chondroitin sulfate biosynthesis. This enzyme functions downstream of xylosyltransferases XYLT1/2 and galactosyltransferase I (B4GALT7), and cooperates with B3GAT3 to complete the linker. Its activity is regulated by upstream factors including TGF-?? signaling, the transcription factor SOX9, and UDP-galactose availability. Loss of B3GALT6 disrupts the assembly of proteoglycans such as syndecans, glypicans, aggrecan, and perlecan, consequently altering glycosylation-dependent signaling cascades and extracellular matrix organization.
In the HT29 background, B3GALT6 knockout provides a powerful tool to dissect the roles of proteoglycans in colorectal cancer progression, where altered glycosylation is a hallmark. This model enables analysis of how B3GALT6 loss impacts epithelial barrier integrity, cell adhesion, migration, and growth factor responses. Moreover, as B3GALT6 mutations are linked to linkeropathies such as spondylodysplastic Ehlers-Danlos syndrome and spondyloepimetaphyseal dysplasia, this cell population serves as a relevant platform for studying disease mechanisms and testing therapeutic interventions in a cancer-relevant context.
This polyclonal knockout population is suitable for a range of applications, including investigations of GAG biosynthesis, drug screening targeting glycosylation pathways, and mechanistic studies of proteoglycan-mediated signaling. Representative assays comprise Western blotting for proteoglycan core proteins, RT-qPCR of glycosyltransferase genes, flow cytometry with lectins to assess cell surface glycans, migration and invasion assays, and barrier integrity measurements. RNA-seq and metabolic flux analysis of sugar nucleotides can further characterize transcriptomic and metabolic reprogramming upon B3GALT6 loss. For detailed technical specifications, validation data, and ordering information, please contact Ascent Research.
