The CGN Knockout Raji Polyclonal Cells are a CRISPR/Cas9-mediated polyclonal knockout cell population derived from the human Raji B lymphocyte cell line, specifically engineered to disrupt the CGN gene, which encodes the tight junction scaffolding protein cingulin. This pooled format provides a heterogeneous loss-of-function model suitable for functional genomics studies without requiring clonal expansion, allowing robust assessment of gene function across a diverse cell population.
The parental Raji cell line is an Epstein-Barr virus (EBV)-positive Burkitt??s lymphoma line that expresses hallmark B cell surface markers including CD19 and CD20, making it a standard model for B cell immunology and lymphoma research. As a suspension cell line, Raji offers a physiologically relevant non-epithelial background to investigate cingulin??s cellular roles beyond its well-characterized function in tight junction assembly and epithelial barrier maintenance.
Cingulin is a central tight junction plaque protein that physically connects transmembrane adhesion receptors??such as occludin, claudins, and JAM-A??to the actin cytoskeleton by binding zonula occludens proteins ZO-1 and ZO-2, as well as myosin II. Critically, cingulin regulates RhoA signaling through direct interaction with the guanine nucleotide exchange factor GEF-H1 (ARHGEF2), thereby controlling actin polymerization and paracellular permeability. This signaling node is modulated upstream by the small GTPases RhoA and Rac1, myosin light chain kinase (MLCK), and protein kinase C (PKC), placing cingulin at a convergence point for cytoskeletal and adhesion-driven signals.
In the Raji lymphocyte context, cingulin’s function is largely unexplored, yet evidence suggests its involvement in processes relevant to lymphoma progression, such as cell adhesion, migration, and RhoA-dependent cytoskeletal reorganization. This knockout model enables the dissection of cingulin-dependent signaling pathways in B cell biology, potentially uncovering roles in immune cell activation, trafficking, or oncogenic transformation. Moreover, it serves as a valuable tool for studying the contribution of tight junction-associated proteins to cancer metastasis, given the known link between junctional proteins and tumor cell dissemination.
This product supports a broad range of research applications, including tight junction biology, paracellular permeability assays (such as TEER measurement and paracellular flux assays), and high-throughput screening for tight junction modulators. Typical experimental readouts include Western blotting and RT-qPCR for knockout confirmation, immunofluorescence and co-immunoprecipitation for protein interaction analysis, RhoA activation assays to probe signaling, and actin staining and cell adhesion assays to assess cytoskeletal effects. The polyclonal nature makes these cells well-suited for pooled CRISPR screens and parallel functional assays. For additional information, please contact Ascent Research.
