The CENPV Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Raji B lymphocyte line. This product provides a genetically heterogeneous pool of cells with targeted disruption of the CENPV gene, enabling loss-of-function studies in a Burkitt lymphoma background. The polyclonal format preserves the diversity of editing events across the population, which is useful for assessing general gene knockout effects without clonal selection bias. These cells serve as a versatile tool for investigating centromere biology and mitotic regulation.
Raji cells are a well-characterized human Burkitt lymphoma cell line isolated from an 11-year-old male. They are EBV-positive and grow in suspension, displaying characteristic B-cell markers such as CD19, CD20, and surface IgM. These cells are widely utilized as a model system for B-cell malignancies, particularly for studying lymphomagenesis, antibody production, and immune response. Their rapid proliferation and susceptibility to mitotic disruptions make them an ideal host for examining genes involved in chromosome segregation.
CENPV encodes a constitutive centromere protein essential for kinetochore assembly and proper chromosome segregation. The protein functions within the centromere/kinetochore complex, interacting with CENP-A, CENP-B, CENP-C, CENP-H, CENP-I, CENP-K, and CENP-M, and facilitating microtubule attachment via the NDC80 complex. CENPV is transcriptionally regulated by E2F factors and loaded in a cell cycle-dependent manner. Its disruption impairs mitotic checkpoint signaling by affecting downstream targets such as BUB1, BUBR1, and MAD2, leading to chromosome misalignment, anaphase bridges, and aneuploidy.
In the context of Raji lymphoma cells, CENPV knockout is particularly consequential. Burkitt lymphoma cells exhibit high mitotic rates and inherent chromosomal instability; loss of CENPV exacerbates these defects, precipitating severe mitotic catastrophe and cell death. This model allows dissection of kinetochore dysfunction specifically in B-cell neoplasia and aids in evaluating therapeutic strategies that target the mitotic machinery. The polyclonal population captures a range of knockout effects, offering a realistic simulation of heterogeneous tumor cell responses.
Researchers can employ these knockout cells in diverse applications, including investigation of centromere identity, kinetochore assembly, and the spindle assembly checkpoint. Typical assays include immunofluorescence staining for centromere/kinetochore markers, Western blotting of CENPV and associated proteins, flow cytometric cell cycle analysis, and assessment of chromosomal instability via micronucleus scoring. Furthermore, they are suitable for drug sensitivity profiling with microtubule poisons or other mitotic inhibitors, RNA-seq transcriptional profiling, and apoptosis studies. For further information, please contact Ascent Research.
