NECTIN2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the NECTIN2 gene in the Raji human B lymphoblastoid cell line. This polyclonal knockout model introduces target-gene disruption across a heterogeneous pool of edited cells, enabling functional studies of NECTIN2-dependent processes without the selection bias associated with clonal isolates. The product is designed to serve as a loss-of-function model for investigating NECTIN2 biology in immune checkpoint regulation, cell adhesion, and viral entry, and is suitable for a broad range of downstream assays including flow cytometry, functional cytotoxicity analyses, and gene expression profiling.
The Raji cell line is a well-characterized human B lymphocyte model derived from a Burkitt lymphoma patient and is persistently infected with Epstein-Barr virus. These cells exhibit a lymphoblastoid morphology, express B cell surface markers, and retain key features of antigen-presenting cells, including the ability to form immune synapses. Raji cells are widely employed in immunology and oncology research to study B cell biology, lymphomagenesis, and tumor-immune interactions. Their robust growth in suspension culture, ease of genetic manipulation, and well-documented signaling properties make them an ideal host for generating CRISPR/Cas9-edited knockout populations aimed at dissecting molecular mechanisms underlying immune recognition and evasion.
NECTIN2, also known as CD112 or poliovirus receptor-related 2, is a transmembrane cell adhesion molecule belonging to the immunoglobulin superfamily. It mediates both homophilic interactions with itself and heterophilic binding with NECTIN3, contributing to intercellular junction formation. Importantly, NECTIN2 functions as a dual ligand for the activating receptor DNAM-1 (CD226) and the inhibitory receptors TIGIT and CD96 on natural killer cells and T cells. This interaction network orchestrates the balance between immune activation and suppression, with NECTIN2?CDNAM-1 binding promoting NK cell cytotoxicity and T cell priming, while NECTIN2?CTIGIT ligation delivers inhibitory signals that dampen immune responses. Downstream of NECTIN2 engagement, signaling pathways involving RAC1, CDC42, and the adaptor protein Afadin regulate cytoskeletal reorganization and cell adhesion dynamics. The expression of NECTIN2 is modulated by upstream stimuli such as interferon-gamma, tumor necrosis factor, and Toll-like receptor ligands, linking it to inflammatory microenvironments.
In the Raji B lymphoma background, knockout of NECTIN2 disrupts the balance of activating and inhibitory signals that modulate interactions with cytotoxic lymphocytes. Because Raji cells naturally express NECTIN2, they can engage DNAM-1 and TIGIT on NK cells and T cells, thereby influencing susceptibility to immune-mediated killing. Loss of NECTIN2 in these cells eliminates this dual signaling platform, enabling researchers to dissect how the DNAM-1/TIGIT axis controls lymphoma cell recognition and clearance. This model is particularly valuable for studying immune evasion mechanisms in Burkitt lymphoma and other B cell malignancies, where NECTIN2 overexpression may contribute to the suppression of antitumor immunity through TIGIT-mediated inhibition. Additionally, since Raji cells harbor EBV, the knockout may alter interactions with viral proteins, as NECTIN2 has been implicated in herpes simplex virus glycoprotein D binding.
Typical research applications of NECTIN2 Knockout Raji Polyclonal Cells include cancer immunotherapy target validation, exploration of immune checkpoint biology, lymphoma functional studies, and investigation of herpesvirus entry mechanisms. The polyclonal knockout population can be characterized using flow cytometry for surface NECTIN2 expression, Western blotting, and RT-qPCR to confirm gene disruption. Functional assays such as NK cell cytotoxicity assays, T cell activation assays, and TIGIT binding assays are directly enabled by this model. Cell adhesion and aggregation assays can further probe NECTIN2-dependent intercellular interactions. Additionally, viral infection assays with herpes simplex virus can be employed to examine the role of NECTIN2 in viral attachment. For additional information or technical support, please contact Ascent Research.
