The MANBA Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of human B lymphocytes, establishing a loss-of-function model for the MANBA gene. This product provides a heterogeneous pool of edited Raji cells, avoiding clonal selection bias, and is designed for investigating lysosomal beta-mannosidase function. The polyclonal format reflects the genetic diversity of the edited pool, enabling robust population-level analyses of N-glycan degradation processes.
The Raji host cell line is derived from Burkitt??s lymphoma and is EBV-positive, exhibiting B lymphocyte properties including antibody secretion, antigen presentation, and immunological memory. Its active glycoprotein processing and established use in immune research make it well-suited for studying lysosomal enzyme defects in a B-cell context, with consistent growth and assay compatibility.
MANBA encodes lysosomal beta-mannosidase, the terminal enzyme in N-glycan degradation that removes beta-linked mannose residues from glycoproteins. It operates within a pathway including MAN2B1, FUCA1, and HEXA, and is transcriptionally regulated by TFEB and TFE3, master regulators of lysosomal gene expression. MANBA activity yields free mannose and truncated N-glycans, and it functionally interacts with LAMP1 in the lysosomal membrane. Loss of MANBA results in accumulation of mannose-rich oligosaccharides, disrupting lysosomal catabolism and downstream cellular signaling, thereby modeling the molecular pathology of beta-mannosidosis.
In Raji B cells, MANBA knockout provides a disease-relevant model to dissect lysosomal storage disorder mechanisms within professional antigen-presenting cells. This system facilitates exploration of how impaired N-glycan breakdown alters antibody glycosylation, antigen processing, and immune memory functions. It also enables studies connecting lysosomal stress to B-cell survival and activation, offering a platform to investigate the interplay between glycobiology and adaptive immunity in the context of beta-mannosidosis.
Researchers can employ this polyclonal knockout in diverse assays, including western blotting and RT-qPCR for gene expression analysis, immunofluorescence to track LAMP1 localization, and fluorogenic enzymatic assays for quantifying beta-mannosidase activity. Mass spectrometry-based glycan profiling reveals substrate accumulation, while flow cytometry assesses lysosomal mass changes and apoptosis induction. These applications support mechanism-of-action studies, drug screening for lysosomal function enhancers, and exploration of TFEB/TFE3 pathway modulators. For further information, please contact Ascent Research.
