The GALM Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Raji B lymphocyte cell line. This polyclonal pool carries CRISPR/Cas9-mediated disruption of the GALM gene, which encodes galactose mutarotase, an enzyme critical for galactose metabolism. The polyclonal format provides a heterogeneous population of GALM-disrupted cells, enabling robust functional studies without clonal selection biases. This knockout model serves as a valuable tool for investigating galactose utilization, glycoconjugate biosynthesis, and metabolic dependencies in lymphoblastoid cells.
The Raji cell line was established from a Burkitt lymphoma patient and remains Epstein-Barr virus (EBV)-positive. Raji cells grow in suspension as lymphoblasts and exhibit characteristics of mature B lymphocytes, including antibody production and antigen presentation. Their active secretion of immunoglobulins and expression of surface glycoproteins make them an ideal model for studying B cell effector functions and glycan-related processes. The EBV-driven proliferation provides a robust background for metabolic investigations, as these cells rely on efficient nutrient utilization and biosynthetic pathways.
Galactose mutarotase (GALM) catalyzes the interconversion of ??- and ??-hexose anomers, providing ??-D-galactose for the Leloir pathway. In this pathway, galactokinase (GALK1), galactose-1-phosphate uridylyltransferase (GALT), and UDP-galactose-4-epimerase (GALE) convert galactose into UDP-glucose and UDP-galactose, critical for glycoconjugate biosynthesis and energy production. Upstream regulators of GALM include HNF1A and insulin signaling, while its product feeds into reactions governed by GALK1 and hexokinase, with sugar uptake mediated by GLUT family transporters. Knockout of GALM disrupts galactose anomeric equilibrium, reducing Leloir pathway flux and impairing glycoprotein, glycolipid, and N-glycan synthesis.
In Raji B lymphocytes, efficient glycosylation is vital for immunoglobulin secretion, MHC-mediated antigen presentation, and co-stimulatory receptor function. GALM disruption is thus poised to compromise glycan-dependent immune processes and may alter cell proliferation, especially in galactose-containing environments. This model offers a system to study the metabolic interplay between carbohydrate flux and B cell effector functions, with relevance to lymphomagenesis and immune regulation.
Applications include galactose mutarotase activity assays, RT-qPCR, and Western blotting for pathway validation. LC-MS metabolomics and proliferation assays in galactose media probe metabolic rewiring. Lectin blotting and flow cytometry assess N-glycosylation changes. These cell pools are valuable for galactosemia modeling, cancer metabolism research, and immune cell glycoengineering. For more information, contact Ascent Research.
