The CEP83 Knockout Raji Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population of Raji B lymphocytes in which the CEP83 gene has been disrupted, yielding a loss-of-function model for the centrosomal protein CEP83. This polyclonal knockout pool retains the inherent genetic heterogeneity of the Raji background while uniformly lacking functional CEP83 expression, making it suitable for studies where clonal variation is not desired and population-level phenotypes are assessed. The knockout was generated using CRISPR/Cas9-mediated targeted gene disruption without selection for specific clonal genotypes, ensuring representation of diverse editing outcomes.
The host Raji cell line is an Epstein-Barr virus (EBV)-positive human B lymphocyte line derived from a Burkitt lymphoma, exhibiting a lymphoblastoid morphology and rapid suspension growth. Raji cells are widely employed in immunological and oncological research due to their well-characterized signaling pathways and ease of culture. Importantly, Raji cells are non-ciliated and do not assemble a primary cilium, providing a distinct context for examining CEP83 functions beyond ciliogenesis.
CEP83 encodes a core distal appendage protein of the mother centriole that scaffolds distal appendage assembly, essential for primary cilium formation and centriole-membrane anchoring. It interacts with CEP164, FBF1, SCLT1, CEP89, TTBK2, and Rabin8 to build the distal appendage complex. Activation is regulated by the Rab11-Rabin8 cascade and TTBK2 kinase in response to serum deprivation. Functionally, CEP83 acts upstream of primary cilium assembly and hedgehog signaling, promoting GLI transcription factor activation. CEP83 also participates in centrosome duplication and cell cycle regulation via links to Wnt signaling. Mutations in CEP83 cause Nephronophthisis-18 and related ciliopathies, including renal-retinal syndromes.
Although Raji cells lack primary cilia, CEP83 expression persists and likely contributes to centrosome integrity, microtubule organization, and mitotic fidelity. Disruption of CEP83 in this lymphoblastoid background may compromise centrosome duplication, leading to aberrant mitotic spindle formation, chromosome segregation errors, and altered cell proliferation, thereby modeling centrosome-related defects in B-cell malignancies. This system offers a unique non-ciliated model to dissect the ciliogenesis-independent roles of distal appendage proteins in cell cycle progression and centrosome homeostasis.
This polyclonal knockout cell population is suitable for centrosome biology investigations, cell cycle analyses, and functional studies of ciliopathy genes in lymphocytes. Researchers can employ Western blotting to assess CEP83 protein levels, immunofluorescence microscopy to visualize centrosome structure, qPCR for transcript quantification, cell proliferation and cell cycle flow cytometry assays, and co-immunoprecipitation to probe protein interactions. The model further supports drug screening for centrosome-associated disorders and examination of hedgehog/Wnt pathway crosstalk. For further technical information, please contact Ascent Research.
