The B3GALNT1 Knockout HT29 Polyclonal Cells constitute a CRISPR/Cas9-mediated gene disruption model in the HT29 colorectal adenocarcinoma cell line, generating a heterogeneous polyclonal population with targeted loss-of-function of the B3GALNT1 gene. This product provides a genetically edited cell system designed to interrogate the roles of globo-series glycosphingolipid biosynthesis in epithelial cell biology and colorectal cancer progression. By ablating B3GALNT1 expression, researchers can assess downstream effects on glycosphingolipid-dependent membrane organization, receptor presentation, and intracellular signaling pathways. The polyclonal format offers a practical and robust tool for functional genomics studies without the selective pressures associated with clonal derivation.
The HT29 parental cell line originates from a human colorectal adenocarcinoma of a 44-year-old female and is widely employed as a model for intestinal epithelial barrier function and mucin secretion, as well as a tumorigenic model for colorectal cancer research. HT29 cells retain several characteristics of differentiated enterocytes, including the capacity for polarization and mucus production, making them particularly relevant for investigating how alterations in cell surface glycoconjugates influence epithelial homeostasis and malignant transformation. This well-characterized background enables the study of B3GALNT1 function in a disease-relevant environment, where glycosylation patterns are known to be critical.
B3GALNT1 encodes a Golgi glycosyltransferase that catalyzes transfer of N-acetylgalactosamine to globotriaosylceramide (Gb3) to form globoside (Gb4), a pivotal step in globo-series glycosphingolipid biosynthesis required for P blood group antigen synthesis. Transcriptionally regulated by SP1, this enzyme operates in a network with B4GALNT1, A4GALT, UGCG, and sphingolipid metabolic enzymes, and its product Gb4 serves as the parvovirus B19 receptor, contributes to membrane microdomain organization, and acts as a precursor for downstream glycosphingolipids. Thus, B3GALNT1 disruption eliminates Gb4 and remodels cell surface glycolipid profiles.
In HT29 colorectal adenocarcinoma cells, B3GALNT1 knockout abrogates globo-series glycosphingolipid synthesis, directly linking the model to colorectal cancer glycosylation alterations. This system permits rigorous investigation of how loss of Gb4 impacts tumor cell adhesion, migration, and signaling events that rely on membrane domain integrity, while also enabling studies on parvovirus B19 receptor biology and P1PK blood group antigen functions in epithelial malignancy.
Researchers can employ this polyclonal knockout population for functional glycobiology studies including glycolipid profiling by mass spectrometry, flow cytometric detection of Gb4, and western blot analysis of pathway components. Cell adhesion, migration, and invasion assays under various extracellular matrix conditions clarify Gb4 roles in tumor malignancy, while drug sensitivity testing may uncover glycosylation-dependent therapeutic responses. Additional applications encompass transcriptomic profiling by RNA-seq and pathogen binding assays for parvovirus B19. For further details, please contact Ascent Research.
