This CRISPR/Cas9-edited polyclonal knockout cell population features targeted disruption of the EXOC6B gene in the Raji B lymphocyte line. EXOC6B encodes a subunit of the exocyst complex, a key tethering factor for post-Golgi vesicles. The polyclonal pool contains cells with heterogeneous indels, offering a robust loss-of-function model without clonal isolation. This format is ideal for experiments requiring genetic diversity or where polyclonal cultures mimic population-level gene inactivation. It provides a reliable system for studying EXOC6B functions in vesicle trafficking and cell polarity within a B cell context.
Raji cells are derived from human Burkitt lymphoma, a highly aggressive B-cell malignancy. This established cell line exhibits characteristics of mature B lymphocytes and is extensively used in immunology and cancer research, particularly for investigating B cell receptor signaling, viral oncogenesis, and apoptosis. As a cancerous B lymphocyte model, Raji cells offer a relevant platform for examining lymphomagenesis and immune cell function. Their rapid growth and ease of genetic modification facilitate efficient generation of knockout polyclonal populations.
EXOC6B is a core subunit of the exocyst complex that mediates tethering of post-Golgi vesicles to the plasma membrane, a pivotal step in targeted exocytosis and polarized secretion. The complex is activated by Ral GTPases downstream of PI3K/AKT and growth factor receptor signaling, and coordinates with Cdc42 and the Par polarity complex. EXOC6B facilitates downstream events including SNARE-mediated fusion, secretion of MMP2/MMP9, and regulation of integrin recycling and GLUT4 translocation. This network underpins cell migration, insulin-stimulated glucose transport, and neurite outgrowth.
Disruption of EXOC6B in Raji polyclonal cells profoundly impairs exocyst-dependent exocytosis, leading to defective post-Golgi vesicle targeting and diminished polarized secretion. In B lymphocytes, the exocyst complex is implicated in the directed release of immunoglobulins and cytokines; thus, this knockout model enables dissection of how exocyst-driven trafficking supports B cell effector functions and malignant transformation. Loss of EXOC6B likely alters the secretion of matrix-remodeling MMPs, disrupts integrin recycling, and attenuates directional migration, processes essential for lymphoma dissemination.
This knockout polyclonal cell population is amenable to a wide array of assays, including Western blotting, immunofluorescence, and flow cytometry for protein expression and cell surface marker profiling. ELISA-based secretion assays can quantify changes in immunoglobulin or MMP release, while Transwell migration and invasion assays directly measure functional consequences on cell motility. Cell polarity analysis via immunofluorescence and transcriptomic profiling by RNA-seq provide deeper mechanistic insights. These approaches collectively enable robust investigation of B cell exocytosis, immune cell polarization, and cancer cell dissemination. For further details, please contact Ascent Research.
