The MAN2C1 Knockout Raji Polyclonal Cells are a heterogeneous population of human B lymphocytes derived from the Raji cell line, engineered via CRISPR/Cas9-mediated gene disruption to eliminate functional expression of MAN2C1 (alpha-mannosidase 2C1). This polyclonal knockout pool is generated without single-cell cloning, yielding a mixed population of edited cells suitable for pooled functional studies, loss-of-function screens, and bulk biochemical analyses. The product enables researchers to interrogate MAN2C1-dependent processes in a lymphoma model without the confounding effects of monoclonal selection or clonal adaptation, providing a physiologically relevant system for studying tumor suppressor regulation and glycobiology.
Raji cells are an Epstein-Barr virus-positive Burkitt lymphoma line originating from a pediatric B cell malignancy. As B lymphocytes, they exhibit features of mature B cells, including surface immunoglobulin expression, antigen presentation capacity, and a proliferative phenotype driven by MYC translocation. These cells serve as a widely used model for B-cell lymphomagenesis, antibody production, and adaptive immunity, offering a relevant context for dissecting oncogenic signaling and lysosomal biology. Their rapid growth and amenability to genetic manipulation make them ideal for generating CRISPR-modified pools for functional genomics and drug response profiling.
MAN2C1 encodes a ubiquitous lysosomal and cytosolic alpha-mannosidase that catalyzes the removal of mannose residues from N-linked glycoproteins, playing a pivotal role in glycoprotein quality control and protein stability. Critically, MAN2C1 de-mannosylates and stabilizes the tumor suppressor PTEN, shielding it from proteasomal degradation. PTEN is a key negative regulator of the PI3K/AKT signaling pathway; its stabilization by MAN2C1 leads to suppression of AKT phosphorylation and downstream mTOR activity, promoting pro-apoptotic signaling via BAX and caspases. MAN2C1 expression is regulated by cellular stress, p53, MYC, and growth factor signaling, positioning it as a node linking glycobiology to tumor suppression. Loss of MAN2C1 disrupts PTEN stability, leading to constitutive AKT activation, enhanced cell survival, and proliferation. This protein network also interfaces with lysosomal proteases and N-linked glycoprotein substrates, highlighting the crosstalk between lysosomal catabolism and signaling.
In the Raji lymphoma background, MAN2C1 knockout models the oncogenic consequences of PTEN loss. Constitutive AKT signaling driven by PTEN degradation promotes unchecked B-cell proliferation, resistance to apoptosis, and lymphomagenesis, mirroring aggressive B-cell malignancies. The absence of MAN2C1 disrupts N-glycan degradation pathways, potentially altering glycoprotein function and membrane dynamics, which may contribute to immune evasion or altered antigen presentation in Burkitt lymphoma. This model is thus a powerful tool for dissecting how post-translational stabilization of tumor suppressors by lysosomal enzymes controls lymphoma biology, providing insights into therapeutic vulnerabilities at the intersection of glycosylation and PI3K/AKT-driven oncogenesis.
Typical applications include mechanistic studies of PTEN regulation, PI3K/AKT pathway activation, apoptosis resistance, and N-glycan processing in B-cell malignancies. The cell pool supports a range of assays: Western blotting to assess PTEN and phospho-AKT levels; flow cytometry for annexin V-based apoptosis detection; cell proliferation measurements via MTS or CFSE; xenograft tumor growth analyses; drug sensitivity screens targeting AKT or mTOR; transcriptomic profiling (RNA-seq); and quantitative N-glycomics. These applications facilitate therapeutic target validation and glycobiology research, positioning the MAN2C1 knockout as a versatile model for cancer biology and drug discovery. For further information or technical support, please contact Ascent Research.
