The B4GALT7 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from HT29 colorectal adenocarcinoma cells, with disruption of the B4GALT7 gene. This heterogeneous knockout model supports studies of glycosaminoglycan biosynthesis and proteoglycan function without clonal selection. The targeted gene disruption impairs galactosyltransferase activity required for initiating glycosaminoglycan chain assembly on core proteins, offering a tool for investigating extracellular matrix biology and cell surface signaling in a physiologically relevant epithelial context.
The HT29 host cell line originates from a primary colorectal adenocarcinoma of a 44-year-old female and serves as an intestinal epithelial model. These cells produce mucin and form polarized monolayers, enabling studies of epithelial barrier function, mucus secretion, and transepithelial transport. Their colorectal tumor origin also makes them relevant for investigating colon cancer progression and the tumor microenvironment.
B4GALT7 encodes a galactosyltransferase that adds galactose to xylose residues in the tetrasaccharide linkage region of proteoglycans, a step essential for heparan sulfate and chondroitin sulfate polymerization. It acts in concert with XYLT1, XYLT2, and B3GALT6, and depends on UDP-galactose and manganese. Downstream, modification of core proteins including syndecans, glypicans, and decorin affects growth factor binding, cell adhesion, and matrix organization. B4GALT7 disruption therefore compromises multiple proteoglycan species and alters glycocalyx and extracellular matrix composition.
In HT29 cells, B4GALT7 loss disrupts cell surface and secreted proteoglycans, potentially affecting colorectal cancer processes such as proliferation, migration, and epithelial-mesenchymal transition. Altered glycosaminoglycan profiles may also impact mucin production and the protective mucus barrier, modulating microbiota interactions and drug absorption. This polyclonal knockout pool mirrors tumor heterogeneity, enabling robust investigation of proteoglycan-dependent signaling in intestinal pathology.
Researchers can apply this B4GALT7 knockout polyclonal cell pool in diverse assays. Functional readouts include cell adhesion and migration assays, transepithelial electrical resistance (TEER) for barrier integrity, and alcian blue staining for sulfated glycosaminoglycans. Molecular approaches such as western blotting for protein glycosylation, RT-qPCR of GAG biosynthesis enzymes, and immunofluorescence or flow cytometry for cell surface proteoglycan expression provide mechanistic detail. HPLC-MS-based disaccharide analysis further allows profiling of glycosaminoglycan composition, supporting studies on drug metabolism and transport in a colorectal epithelial model. For further details, contact Ascent Research.
