The CDK5RAP2 Knockout Raji Polyclonal Cells product consists of a heterogeneous population of Raji cells that have undergone CRISPR/Cas9-mediated disruption of the CDK5RAP2 gene. This polyclonal knockout pool provides a loss-of-function model for studying CDK5RAP2-dependent processes in a human B-lymphocyte context. The gene editing approach generates a mixed population of edited alleles, enabling the investigation of CDK5RAP2 function without the clonal artifacts or selection pressures inherent to single-cell-derived lines. This product is well-suited for functional genomics, phenotypic screening, and pathway dissection experiments.
The parental Raji cell line is a suspension-adapted B lymphocyte derived from a patient with Burkitt lymphoma. These cells are Epstein-Barr virus (EBV)-positive and are widely utilized in immunological research, including studies of B-cell biology, EBV latency and reactivation, and antibody-dependent cell-mediated cytotoxicity (ADCC) assays. Raji cells express key B-cell markers and maintain active signaling pathways relevant to lymphocyte proliferation and survival. Their robust growth in suspension culture and well-characterized genomic landscape make them a reproducible host for gene knockout experiments.
CDK5RAP2 encodes a centrosomal scaffold protein that anchors the ??-tubulin ring complex (??-TuRC) to centrosomes via direct interaction with ??-tubulin (TUBG1) and pericentrin (PCNT), thereby promoting microtubule nucleation and organization. CDK5RAP2 is regulated by upstream kinases including CDK5, PLK1, and Aurora A, which modulate its localization and activity during the cell cycle. It forms critical complexes with CEP152, CEP63, and WDR62 to coordinate centriole duplication and spindle pole integrity. Downstream, CDK5RAP2 controls spindle orientation through the NEDD1?C??-TuRC axis and influences the recruitment of CEP152 and PLK4 for procentriole assembly. Loss of CDK5RAP2 disrupts mitotic spindle alignment, leading to defects in chromosome segregation and asymmetric cell division, which are especially consequential in neural progenitor cells where it is linked to primary microcephaly type 3 (MCPH3) and Seckel syndrome.
In the Raji B-cell context, CDK5RAP2 knockout allows investigation of centrosome-dependent processes within lymphocytes, a cell type where centrosome function is critical for polarized signaling during immune synapse formation and asymmetric cell division during clonal expansion. The EBV-positive background offers a unique opportunity to study virus?Chost interactions at the centrosome, as EBV gene products can perturb mitotic machinery. Additionally, Raji cells are a model for Burkitt lymphoma, and CDK5RAP2 disruption may uncover vulnerabilities in centrosome-amplified tumor cells, enabling exploration of synthetic lethal strategies or centrosome-targeted therapies. This system thus bridges basic centrosome biology with translational oncology and neurodevelopmental disease modeling.
Researchers can employ this knockout model in a broad range of applications, including dissection of mitotic spindle regulation by immunofluorescence staining for ??-tubulin and pericentrin, cell cycle profiling via flow cytometry, and proliferation or apoptosis assays to evaluate downstream phenotypic consequences. RNA sequencing or proteomic analyses of the CDK5RAP2-deficient polyclonal population can reveal transcriptional and signaling network adaptations. Co-immunoprecipitation experiments may dissect altered interactions with core partners such as TUBG1, PCNT, CEP152, or WDR62. Drug-screening campaigns can test compounds targeting centrosome clustering or microtubule dynamics. For further technical details, protocol recommendations, or custom applications, please contact Ascent Research.
