The MGAT5 Knockout Raji Polyclonal Cells product comprises a heterogeneous population of Raji B lymphocytes with CRISPR/Cas9-mediated disruption of the MGAT5 gene, encoding N-acetylglucosaminyltransferase V (GnT-V). This polyclonal knockout cell pool provides a loss-of-function model to study the impact of MGAT5 ablation on glycan branching and downstream cellular processes.
The Raji cell line is an Epstein-Barr virus (EBV)-positive B lymphoblastoid line derived from a Nigerian patient with Burkitt lymphoma. These suspension cells express characteristic B-cell markers including CD19, CD20, and surface IgM, making them a well-established model for B-cell biology, EBV latency, and lymphoma pathogenesis. Raji cells are widely employed to investigate signaling networks governing proliferation, adhesion, and immune evasion in malignant B cells.
MGAT5 encodes GnT-V, a Golgi-resident glycosyltransferase that catalyzes the addition of ??1,6-linked N-acetylglucosamine to the core mannose of N-glycans, thereby generating multi-antennary complex structures on glycoproteins. This branching modification is regulated by upstream factors such as TGF-??, EGF, Ets-1, and c-Jun, and critically affects the stability and function of downstream targets including integrins (??5??1, ??V??3), EGFR, MET, and cadherins. GnT-V-mediated glycosylation of integrins and growth factor receptors enhances their clustering and prolongs signaling through FAK, Src, PI3K-Akt, and ERK pathways, thereby promoting cell adhesion, migration, and tumorigenicity. The enzyme functions in concert with other Golgi glycosyltransferases and uses UDP-GlcNAc as the donor substrate for its catalytic activity.
Disruption of MGAT5 in Raji cells ablates GnT-V activity, leading to a marked reduction in ??1,6-branched N-glycans on the cell surface detectable by L-PHA lectin blotting. This loss impairs integrin- and EGFR-mediated signaling cascades, attenuating FAK, Akt, and ERK phosphorylation and consequently diminishing cell adhesion, spreading, and motility. In the context of B-cell lymphoma, MGAT5 knockout may alter immune synapse formation, natural killer cell recognition, and general glycocalyx organization, providing a powerful tool to dissect how altered glycosylation influences malignant behavior and immune surveillance.
This MGAT5 knockout polyclonal population enables a wide range of downstream functional assays, including quantitative assessment of ??1,6-branched glycans by flow cytometry, adhesion and migration assays, proliferation kinetics, and phospho-signaling analyses via Western blotting. Researchers can employ these cells to validate glycogene targets in cancer biology, investigate glycan-dependent mechanisms of metastasis, explore glycoimmune interactions, and test modulators of glycosylation in drug discovery. RNA-seq and tumor xenograft studies can further elucidate transcriptomic and in vivo consequences of MGAT5 loss. For additional information, including batch-specific data and custom services, please contact Ascent Research.
