The EVI5 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B-lymphocyte cell line. This product comprises a heterogeneous pool of cells with targeted disruption of the EVI5 gene, achieved via CRISPR/Cas9-mediated gene editing. The polyclonal format provides a robust loss-of-function model without clonal selection, enabling the study of EVI5-dependent processes in a genetically mixed background that may better reflect physiological variability. Researchers can use these polyclonal knockout cells to investigate EVI5 function in B-cell lymphoma biology and cell cycle regulation.
The parental Raji cell line is derived from an Epstein-Barr virus (EBV)-positive Burkitt’s lymphoma of an 11-year-old male. Raji cells exhibit a B-lymphoblastoid morphology and serve as a well-characterized model for B-cell malignancies and immune system research. They are widely employed to study oncogenic signaling, apoptosis, and the role of EBV in lymphomagenesis. The availability of EVI5 knockout in this cellular context provides a valuable tool for dissecting EVI5-mediated pathways specifically relevant to aggressive B-cell lymphomas.
EVI5 encodes a Rab GTPase-activating protein (GAP) that specifically regulates Rab35, a small GTPase involved in endocytic recycling and cytokinesis. By stimulating GTP hydrolysis on Rab35, EVI5 modulates trafficking of signaling receptors and adhesion molecules, influencing cell cycle progression at the G1/S transition. EVI5 expression is transcriptionally activated by E2F transcription factors, linking its function to the RB1 pathway. At the centrosome, EVI5 interacts with gamma-tubulin and centrin, contributing to centrosome duplication. Downstream, Rab35 inactivation indirectly promotes cyclin D1 expression, facilitating G1/S transition. The EVI5-Rab35 axis intersects with key cell cycle regulators, including CDK4, cyclin D1, RB1, and E2F1, as well as the endocytic recycling factor EHD1.
In Raji B-lymphoma cells, EVI5 is positioned at a node linking endocytic trafficking to cell cycle entry. Knockout of EVI5 is expected to disrupt Rab35-mediated recycling pathways, leading to aberrant accumulation of surface proteins and altered growth factor signaling. Since Raji cells harbor multiple genetic lesions that accelerate the cell cycle, loss of EVI5 may uncover vulnerabilities in centrosome duplication and cyclin D1-dependent G1 progression. This polyclonal knockout model offers a platform to assess the dependency of Burkitt’s lymphoma cells on EVI5 for proliferation and survival, and to investigate the interplay between Rab GTPase signaling and oncogenic networks in B-cell malignancies.
These polyclonal EVI5 knockout Raji cells are suitable for functional studies. Researchers can employ western blotting or RT-qPCR to confirm EVI5 ablation. Flow cytometry enables cell cycle profiling, and BrdU or MTT assays quantify proliferation. Co-immunoprecipitation and immunofluorescence probe EVI5 interactions with gamma-tubulin or Rab35, while apoptosis assays assess chemosensitivity. This model supports drug target validation in B-cell lymphoma, multiple sclerosis, or neuroblastoma research. For additional details, please contact Ascent Research.
