The EOGT Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Raji B lymphocyte line, featuring targeted disruption of the EOGT gene. This loss-of-function model provides a robust system to study EOGT-mediated O-GlcNAcylation in Notch signaling without clonal selection artifacts.
Raji cells are an EBV?positive Burkitt lymphoma line with suspension lymphoblastoid morphology, routinely used to investigate B?cell receptor signaling, antibody production, and antigen presentation. Their native expression of Notch pathway components makes them highly suitable for examining perturbations in O?GlcNAc?dependent receptor modification. As a model for B?lymphocyte biology, Raji cells endogenously express key elements of the Notch cascade, facilitating direct study of glycosylation?regulated signaling in an immune?relevant background.
EOGT encodes an EGF domain?specific O?GlcNAc transferase that modifies conserved serine and threonine residues within EGF repeats of Notch receptors and ligands, directly impacting their folding, trafficking, and signal transduction. This enzyme functions downstream of ligands such as DLL4 and JAG1 and upstream of NOTCH1 and NOTCH2 receptor cleavage, ultimately modulating RBPJ?dependent transcription of targets like HES1. Key substrates include EGF repeats of NOTCH1, NOTCH2, DLL1, DLL4, JAG1, and JAG2, with UDP?GlcNAc serving as the donor sugar. ER stress has been identified as an upstream regulator capable of altering EOGT activity. EOGT disruption thus abolishes O?GlcNAc modification on these substrates, leading to impaired receptor?ligand complex formation and attenuated downstream gene expression.
In the Raji B?cell context, Notch signaling governs proliferation and survival programs, and its dysregulation is implicated in B?cell lymphomagenesis. The EOGT knockout background allows researchers to isolate the contribution of Notch EGF repeat O?GlcNAcylation to malignant phenotypes without interfering with general O?GlcNAc processing. This model is particularly relevant for exploring links to Adams?Oliver syndrome and for investigating how ligand?specific glycosylation affects B?cell transformation.
Researchers can employ this polyclonal population in Notch luciferase reporter assays, Western blot detection of NOTCH1 cleavage, flow cytometric monitoring of surface Notch expression, and anti?O?GlcNAc immunoprecipitation followed by qPCR for HES1 and MYC. Co?culture activation assays further enable dissection of juxtacrine signaling dynamics in immune cells. These applications position the EOGT knockout cells as a versatile platform for screening O?GlcNAcylation modulators and probing EOGT function in B?cell lymphoma. For additional details, contact Ascent Research.
