The B4GALT3 Knockout U266 Cell Line is a ready-to-use CRISPR/Cas9-edited knockout cell line derived from the human U266 multiple myeloma cell line, featuring targeted disruption of the B4GALT3 gene. This loss-of-function model enables the study of beta-1,4-galactosyltransferase 3 activity in a plasma cell malignancy context without the need for transient knockdown approaches. The cell line is provided as a stable knockout population, allowing for reproducible experiments in glycoscience and myeloma research.
The U266 cell line, originating from a patient with IgE-producing multiple myeloma, serves as a widely used in vitro model for plasma cell biology. These B lymphocytes (plasma cells) express surface CD38 and CD138, secrete immunoglobulin E, and recapitulate key features of malignant plasma cell disorders. Their well-characterized glycoprotein expression profiles and adhesion properties make them a relevant host for studying glycosylation-dependent processes in myeloma.
B4GALT3 encodes a ??1,4-galactosyltransferase localized to the Golgi apparatus, which catalyzes the transfer of galactose from UDP-galactose to terminal N-acetylglucosamine residues on N- and O-linked glycoconjugates. This enzyme is transcriptionally regulated by SP1 and NF-??B in response to upstream cytokines such as IL-6 and TNF-??, and growth factors including EGF and FGF. It functions downstream of these signals to generate ??1,4-galactosyl linkages essential for the synthesis of complex glycans, including sialyl Lewis X structures, and for the modification of adhesion molecules like CD44 and integrins. B4GALT3 interacts with UDP-galactose, N-acetylglucosaminyltransferases, and Golgi matrix proteins to coordinate glycan extension, ultimately influencing selectin ligand biosynthesis and cell-cell adhesion.
Disruption of B4GALT3 in U266 myeloma cells is anticipated to impair terminal galactosylation of glycoconjugates, leading to altered surface expression of glycosylated receptors and adhesion molecules. This model is particularly relevant for multiple myeloma research, as aberrant glycosylation patterns are associated with tumor progression, immune evasion, and drug resistance. The U266 background provides a clinically pertinent system to dissect how galactosylation defects affect IgE secretion, CD38/CD138 surface topology, and interactions with the bone marrow microenvironment, including selectin-mediated adhesion.
Key research applications include glycomic profiling of myeloma cells, investigation of sialyl Lewis X and selectin ligand biosynthesis, and functional studies of adhesion-mediated drug resistance. Researchers can employ flow cytometry with anti-sialyl Lewis X antibodies, lectin blotting using L-PHA and E-PHA, and adhesion assays to E-selectin to characterize glycosylation changes. Migration assays, Western blotting for CD44, RT-qPCR analysis of glyco-gene expression, and glycomic mass spectrometry further enable detailed mechanistic dissection. For inquiries regarding this product, please contact Ascent Research.
