The GBA2 Knockout Raji Polyclonal Cells comprise a CRISPR/Cas9-engineered population of Raji B lymphocytes with targeted disruption of the GBA2 gene. This polyclonal knockout pool provides a robust loss-of-function model for investigating non-lysosomal glucosylceramidase activity, enabling direct interrogation of GBA2-dependent sphingolipid metabolic pathways without the confounding presence of wild-type alleles in the bulk cell population. The cells are delivered as a ready-to-use suspension culture suitable for downstream phenotypic, biochemical, and pharmacological analyses.
The Raji host cell line is a human B lymphoblastoid line derived from Burkitt??s lymphoma. These EBV-positive cells retain immunoglobulin expression and antigen-presentation capacity, and they proliferate rapidly in suspension culture. Widely used in immunology and hematological malignancy research, their well-characterized nature also makes them valuable for lipid membrane trafficking studies, especially given the relevance of glycosphingolipids to B-cell receptor signaling and lipid raft biology.
GBA2 encodes a non-lysosomal glucosylceramidase that hydrolyzes glucosylceramide into ceramide and glucose at the cytosolic face of membranes, operating alongside the lysosomal acid ??-glucosidase GBA1. The enzyme is regulated by ceramide levels, glucosylceramide accumulation, inflammatory cytokines, and lipid stress signals. Its activity impinges on downstream processes including ceramide-mediated apoptosis, mitochondrial function, and autophagy flux. GBA2 cooperates with molecular partners such as ceramide synthase, sphingomyelin synthase, and saposin C, and is embedded in a network of sphingolipid metabolites??ceramide, glucosylceramide, sphingomyelin, acid/neutral sphingomyelinases, ceramide-1-phosphate, and sphingosine-1-phosphate??that collectively dictate cell fate. Consequently, GBA2 disruption deranges this signaling and metabolic equilibrium.
In the Raji B-cell context, GBA2 deletion results in a unique model where non-lysosomal glucosylceramide cleavage is eliminated, causing substrate accumulation and redirection of ceramide flux. This metabolic alteration directly impacts plasma membrane lipid raft composition, potentially modulating B-cell receptor clustering, antigen presentation efficiency, and downstream proliferative or apoptotic responses. Because EBV-driven lymphomagenesis involves sphingolipid-mediated survival pathways, the GBA2 polyclonal knockout offers a physiologically relevant system to dissect how glucosylceramide and ceramide species influence lymphoma cell biology. Moreover, the interplay between GBA2 and the related GBA1 enzyme, which is deficient in Gaucher disease, can be systematically explored in this immune cell background to reveal compensatory or synergistic mechanisms.
This polyclonal knockout product is suited for lipidomic profiling via mass spectrometry to track glucosylceramide and ceramide species, glucosylceramidase activity assays, and quantitative western blotting for sphingolipid enzymes. Flow cytometry can detect ceramide-mediated apoptosis, fluorescence microscopy can visualize lipid distribution, and RT-qPCR can assess metabolic gene expression. Such applications support drug target validation, lysosomal storage disorder modeling, and B-cell malignancy research. For technical details, contact Ascent Research.
