The B4GALT1 Knockout HT29 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the B4GALT1 gene in the human HT29 colorectal adenocarcinoma cell line. This loss-of-function model is designed to investigate the roles of beta-1,4-galactosyltransferase 1 in lactosamine biosynthesis and glycoconjugate-dependent cellular functions.
HT29 cells are derived from a primary colorectal adenocarcinoma in a 44-year-old female and display adherent epithelial morphology with well-characterized APC and TP53 tumor suppressor gene mutations. Under appropriate conditions, they can undergo differentiation into enterocyte-like cells, providing a robust system for studying colonic epithelial biology, oncogenic Wnt signaling, and intestinal cell maturation.
B4GALT1 encodes a Golgi-resident glycosyltransferase that catalyzes the transfer of galactose from UDP-galactose to terminal N-acetylglucosamine (GlcNAc) residues, forming Gal??1-4GlcNAc (lactosamine) on glycoproteins and glycolipids. Its regulation involves butyrate, phorbol esters (PMA), prolactin, and the Wnt/??-catenin pathway, and it functionally interacts with alpha-lactalbumin to modify substrate specificity. The lactosamine structures generated by B4GALT1 are essential for the glycosylation of integrins, E-cadherin, EGFR, and MUC1 mucin, thereby influencing cell adhesion, migration, and intracellular signaling. Knockout of B4GALT1 abolishes poly-N-acetyllactosamine chain synthesis and alters galectin-binding epitopes, leading to impaired cell?Cmatrix and cell?Ccell interactions.
In HT29 cells, B4GALT1 disruption provides a powerful platform to examine glycosylation-dependent mechanisms within the colorectal adenocarcinoma setting, where APC and TP53 mutations drive oncogenesis. This polyclonal model enables interrogation of how lactosamine deficiency affects tumor cell adhesion, migration, and invasiveness, as well as the interplay between glycosylation and epithelial differentiation. It is also relevant for studying congenital disorders of glycosylation type IId and the contribution of altered glycans to cancer metastasis.
Typical experimental uses include lectin blotting with RCA?I to assess galactosylation, flow cytometry for lactosamine epitope detection, cell adhesion assays on ECM proteins, and migration/invasion Transwell assays. N?glycan mass spectrometry profiling, RT?qPCR for glycosylation-related genes, and immunofluorescence for glycoconjugate localization are additional applications. This polyclonal knockout population is also suitable for evaluating galectin?Cglycan interactions and their functional consequences. For further information, please contact Ascent Research.
