The MAPRE2 Knockout Raji Polyclonal Cells represent a genetically engineered population of Raji B lymphocytes in which the MAPRE2 gene has been disrupted via CRISPR/Cas9-mediated genome editing. This polyclonal knockout pool provides a heterogeneous loss-of-function model suitable for investigating microtubule cytoskeletal dynamics in a lymphoblastoid background. By targeting MAPRE2, which encodes the microtubule plus-end tracking protein EB2, this model enables systematic exploration of microtubule regulation in B-cell biology without the constraints of single-clone artifacts.
The Raji cell line, established from a Burkitt lymphoma patient, is an Epstein-Barr virus (EBV)-positive lymphoblastoid cell line that grows in suspension and retains key features of B lymphocytes. Widely utilized as a model for Burkitt lymphoma and B-cell immunology, Raji cells offer a relevant platform for studying oncogenic processes and lymphocyte signaling. Their transformed phenotype and continuous proliferation make them amenable to genetic manipulation and high-throughput functional assays, enabling dissection of pathways that depend on microtubule dynamics during cell division and interphase.
MAPRE2 (EB2) functions as a core plus-end tracking protein (+TIP) that stabilizes microtubules by recruiting CLIP-170 and p150Glued (DCTN1) to growing ends. Interaction with EB1 (MAPRE1) and APC positions MAPRE2 at the center of a regulatory hub, where upstream kinases Aurora A, GSK3??, and CDK1 modulate its activity to control mitotic spindle orientation and cell migration. Disruption of MAPRE2 disrupts the hierarchical assembly of +TIP complexes, compromising downstream microtubule stabilization, cargo transport, and dynein anchoring. In the broader signaling network, MAPRE2 integrates Wnt pathway cues and influences neurodevelopmental processes, highlighting its importance beyond cell autonomous cytoskeletal dynamics.
Within Raji B lymphocytes, MAPRE2-dependent microtubule organization is essential for precise mitotic spindle assembly and chromosome segregation, processes that are frequently dysregulated in lymphoma. EB2 also supports the microtubule remodeling required for immune synapse formation, impacting B-cell activation and antigen presentation. Knocking out MAPRE2 in this lymphoblastoid line thus provides a model to probe how loss of +TIP function contributes to mitotic abnormalities and impaired immune cell interactions, with potential relevance to Burkitt lymphoma pathogenesis and metastasis.
This polyclonal knockout cell population is compatible with immunofluorescence staining of ??-tubulin to assess microtubule network integrity, live-cell imaging of EB1 dynamics to measure plus-end tracking velocity, and western blotting to confirm MAPRE2 protein ablation. Flow cytometry facilitates cell cycle profiling to identify mitotic delays, while transwell migration assays quantify changes in motility. Co-immunoprecipitation enables mapping of altered protein interactions within the +TIP complex. These approaches support research into microtubule-driven regulation of lymphoma cell division, cancer invasion, and neurodevelopmental signaling pathways. For further information, please contact Ascent Research.
