The ERGIC2 Knockout Raji Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ERGIC2 gene in the Raji human B lymphocyte line. This polyclonal format offers a pooled loss-of-function model in which heterogeneous gene disruptions collectively impair ERGIC2 protein function, enabling robust assessment of ER-Golgi intermediate compartment biology without the confounding effects of clonal selection.
Raji cells are a widely used B lymphoblast line, originally derived from an Epstein-Barr virus (EBV)-positive Burkitt lymphoma. These cells retain key features of mature B lymphocytes, including robust antibody secretion, antigen presentation via MHC class II, and activation of adaptive immune signaling pathways. Their high proliferative capacity and genetic manipulability make Raji cells particularly well suited for studying the intersection of secretory pathway regulation and immune cell function.
ERGIC2 encodes an integral membrane protein that serves as a structural scaffold within the ER-Golgi intermediate compartment, where it orchestrates bidirectional vesicular trafficking between the ER and the Golgi apparatus. ERGIC2 forms complexes with ERGIC1 and ERGIC3 and interacts directly with COPII coat components including the SEC23/SEC24 subcomplex and the cargo receptor SURF4, as well as the ER-Golgi SNARE protein SEC22B. These interactions are critical for COPII vesicle formation, which is initiated by SAR1B activation and regulated by the ARF1 GTPase cycle. Downstream, ERGIC2-dependent trafficking governs the anterograde transport of secretory cargo such as immunoglobulins and cell surface glycoproteins, while also facilitating COPI-mediated retrograde retrieval. Disruption of ERGIC2 therefore compromises the entire ER-Golgi transport axis, leading to cargo accumulation, impaired secretion, and defective N-glycosylation of glycoproteins.
Within Raji B cells, ERGIC2 function is particularly consequential due to the high biosynthetic demand for immunoglobulin production and surface expression of the glycosylated B cell receptor. Knockout of ERGIC2 in these lymphoblasts is predicted to diminish antibody secretion and reduce BCR surface levels, thereby perturbing both humoral effector functions and antigen recognition. Moreover, because Raji cells originate from an EBV-associated lymphoma, this model offers a unique platform to examine how secretory pathway dependencies influence lymphomagenesis and immune evasion. The disrupted glycosylation fidelity further mimics aspects of congenital disorders of glycosylation linked to ERGIC trafficking defects, providing a cellular system for dissecting disease mechanisms.
This knockout cell model supports diverse experimental workflows, including live-cell imaging of ER-to-Golgi transport using the VSVG-GFP reporter, biochemical analysis of glycoprotein maturation via lectin blotting, and flow cytometric quantification of surface glycoprotein levels. ELISA and western blotting enable measurement of secreted antibody titers, while immunofluorescence and electron microscopy can localize cargo accumulation within the ER. These applications make the product valuable for investigating B cell biology, secretory pathway vulnerabilities in lymphoma, and the molecular basis of glycosylation disorders. For additional information, please contact Ascent Research.
