NEK9 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in the Raji B lymphocyte background, engineered to abrogate NEK9 gene expression. This polyclonal pool arises from bulk gene disruption and selection, providing a heterogeneous loss-of-function model without the artefacts of clonal isolation. The product is intended for researchers investigating NEK9-dependent biological processes, including mitotic regulation and oncogenic signaling, in a well-characterized human lymphoblastoid system.
The parental Raji cell line was originally derived from a Burkitt lymphoma patient and maintains an Epstein?CBarr virus (EBV)-positive, lymphoblast-like phenotype. Raji cells grow in suspension and serve as a widely used model for B cell biology, humoral immunity, and B-cell malignancies. Their transformed nature and robust proliferation make them particularly amenable to functional genomics studies and high-content screening applications.
NEK9 (NIMA-related kinase 9) is a serine/threonine kinase that functions as a critical regulator of mitotic entry, centrosome separation, and spindle assembly. During the G2/M transition, PLK1 phosphorylates and activates NEK9, which in turn phosphorylates NEK6 and NEK7 to orchestrate centrosome splitting and bipolar spindle formation. Additional upstream regulators include CDK1, AURKA, and the DNA damage-responsive ATM/ATR kinases, while downstream effectors encompass histone H3, BICD2, and LMNA. NEK9 also interacts with NUP98 and CDK5RAP3, integrating mitotic progression with TP53 pathway signaling and cell cycle checkpoints.
In the Raji B-cell lymphoma context, disruption of NEK9 uncouples mitotic control, leading to defects in centrosome separation, aberrant spindle assembly, and ultimately G2/M arrest and apoptosis. This knockout model thus enables dissection of NEK9??s role in lymphoma pathogenesis and DNA damage response, given the upstream ATM/ATR link. It also provides a platform to study how deregulated mitotic signaling cooperates with EBV-driven transformation in B-cell malignancies.
Key applications include investigating mitotic regulation, validating NEK9 as a therapeutic target in B-cell and other NEK9-overexpressing cancers, and performing DNA damage checkpoint analyses. Representative assays suited to these cells are Western blotting for NEK9 and its phosphorylated substrates (e.g., phospho-NEK6, phospho-NEK7), cell cycle profiling by flow cytometry, mitotic index determination, annexin V apoptosis assays, proliferation measurements, immunofluorescence staining of the mitotic spindle, colony formation assays, and transcriptomic profiling via RNA?seq. For further technical information, please contact Ascent Research.
