The Cdc20 Knockout Raji Polyclonal Cells are a heterogeneous population of Raji B lymphocytes engineered through CRISPR/Cas9-mediated disruption of the CDC20 gene. This polyclonal knockout product bypasses clonal isolation, providing a pool of cells harboring diverse loss-of-function mutations. It serves as a robust model for studying the average impact of CDC20 ablation on cell cycle progression and mitotic regulation, without the selective pressures of single-cell cloning, making it ideal for initial functional screens or population-based analyses.
Raji cells originate from a Burkitt??s lymphoma patient and represent an EBV-positive B-lymphoblastoid line. They express characteristic B-cell markers such as CD19, CD20, and CD22, and are widely used in immunology and cancer research. Raji cells grow in suspension with a doubling time of approximately 24 hours, providing a convenient and reproducible host for gene-editing experiments. Their cancerous origin makes them particularly relevant for investigating oncogenic pathways and drug responses in the context of aggressive B-cell lymphoma.
CDC20 is a critical co-activator of the anaphase-promoting complex/cyclosome (APC/C), a multi-subunit E3 ubiquitin ligase governing the metaphase-to-anaphase transition. During early mitosis, the spindle assembly checkpoint (SAC) proteins MAD2, BUBR1, and BUB3 sequester CDC20, preventing premature APC/C activation until proper chromosome attachment. Upon SAC satisfaction, CDC20 integrates into APC/C. Active APC/CCDC20 polyubiquitinates securin and cyclin B, targeting them for proteasomal degradation. This liberates separase for cohesin cleavage and sister chromatid separation, and inactivates CDK1 for mitotic exit. Additionally, CDC20 facilitates timely degradation of Nek2A, contributing to mitotic fidelity.
In Raji cells, disruption of CDC20 is anticipated to impair APC/C function, leading to mitotic arrest, chromosomal missegregation, and apoptosis. Given the oncogenic stress inherent in Burkitt lymphoma??often driven by c-MYC translocation??compromising mitotic regulation may expose synthetic lethal interactions or therapeutic vulnerabilities. This polyclonal knockout model enables detailed dissection of how lymphoma cells cope with mitotic checkpoint dysfunction and provides a platform for evaluating agents that target mitotic machinery.
The Cdc20 Knockout Raji Polyclonal Cells support a broad spectrum of functional assays. Flow cytometry with DNA content staining permits cell cycle profiling after synchronization or treatment with microtubule poisons like nocodazole or taxol. Western blotting and co-immunoprecipitation monitor securin, cyclin B, and Nek2A turnover and APC/C subunit association. Ubiquitination assays measure APC/CCDC20 activity, while confocal microscopy reveals chromosome alignment and spindle defects. These applications facilitate mitotic checkpoint studies, drug sensitivity screens, and functional genomics in lymphoma. For further details, contact Ascent Research.
