The MAN2A1 Knockout Raji Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population designed for investigating N-glycosylation pathways in B lymphocyte biology. This genetically disrupted pool features targeted disruption of the MAN2A1 gene in the Raji B lymphoblastoid cell line, providing a heterogeneous population of cells with ablated Golgi alpha-mannosidase II function. Unlike clonal isolates, this polyclonal format maintains natural response variation while eliminating MAN2A1 activity, making it suitable for experiments requiring population-level glycosylation phenotyping. Researchers can employ this model to study glycan-dependent signaling, adhesion, and immune recognition in a lymphoma context without the bottleneck effects of single-cell cloning.
The Raji host cell line is an EBV-immortalized B lymphoblastoid model originally derived from a Burkitt’s lymphoma patient and carries the hallmark t(8;14) translocation that dysregulates c-MYC expression. These cells are surface immunoglobulin-negative, representing a mature B cell stage arrested prior to antigen receptor expression, and are widely used for studying B cell signaling, EBV latency, and lymphomagenesis. The combination of MAN2A1 knockout with this well-characterized lymphoma background creates a unique system to explore how aberrant N-glycan processing intersects with oncogenic signaling pathways and B cell function.
MAN2A1 encodes Golgi alpha-mannosidase II, a key enzyme in the N-glycan maturation pathway that trims mannose residues to enable the synthesis of complex-type glycans on glycoproteins. This enzyme functions downstream of MGAT1 and upstream of MGAT2, B4GALT1, and ST6GAL1 in the glycosylation cascade. MAN2A1 activity is regulated by unfolded protein response factors such as XBP1 and ATF6, and its expression is influenced by B cell activation signals through CD40 and cytokine receptors. The protein interacts with COPI coatomer complex and Golgi matrix proteins including GM130 and GRASP65 to localize and process substrate glycoproteins, ultimately impacting the glycosylation of cell surface receptors like BCR components and CD45, as well as secreted immunoglobulins.
In the Raji B lymphocyte context, MAN2A1 disruption leads to accumulation of high-mannose glycans on cell surface proteins, impairing B cell receptor signaling, integrin-mediated adhesion, and immune recognition mechanisms. This knockout model recapitulates features of congenital disorders of glycosylation type II (MAN2A1-CDG) and provides insights into how dysregulated glycosylation contributes to lymphoma progression and immune evasion. The interplay between MAN2A1 deficiency and the c-MYC-driven proliferative background of Raji cells enables dissection of glyco-immune checkpoints and the role of protein glycosylation in B cell malignancies.
Typical research applications include studying N-glycan-dependent B cell receptor signaling using phospho-flow cytometry, validating glycosylation-targeting therapeutics via lectin blotting with ConA or L-PHA, and modeling glycosylation disorders through mass spectrometry glycomics. Additional uses encompass examining adhesion molecule function in lymphoma dissemination and investigating how high-mannose glycans alter CD45 and integrin activity. The polyclonal nature allows assessment of glycosylation variability within a population. For further details or to discuss custom applications, please contact Ascent Research.
