The NEK4 Knockout Raji Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population featuring targeted disruption of the NEK4 gene in the human Raji B lymphocyte line. This loss-of-function model is generated via CRISPR/Cas9-mediated gene disruption, resulting in a heterogeneous polyclonal population lacking functional NEK4 kinase activity, without clonal selection. It serves as an essential tool for investigating NEK4-dependent signaling in B-cell biology and lymphoma.
Raji cells are an Epstein-Barr virus (EBV)-positive Burkitt lymphoma-derived B lymphoblastoid line that expresses surface IgM and other B-cell markers, reflecting a mature B-cell phenotype. Widely employed in hematological malignancy research, Raji cells provide a physiologically relevant background for studying B-cell lymphoma biology, DNA damage responses, and apoptosis regulation. Their robust proliferation and well-characterized signaling networks make them suitable for gene-editing approaches to dissect oncogenic and tumor-suppressive pathways.
NEK4 encodes a serine/threonine kinase that integrates multiple cellular processes including ciliogenesis, microtubule dynamics, DNA damage repair, and cell cycle progression. It functions downstream of ATM and ATR kinases in response to DNA damage, phosphorylating and interacting with p53 to modulate apoptosis. NEK4 also participates in Hedgehog and Wnt signaling, where it influences ??-catenin stability and GLI transcription factor activity. Representative pathway components include SMO, FZD, DVL, CHK1, CHK2, BAX, and BCL-2, highlighting NEK4??s role at the intersection of genomic stability and developmental signaling.
In Raji B cells, NEK4 knockout disrupts its kinase activity, impairing DNA damage repair through the ATM/ATR-p53 axis and potentially derepressing p53-mediated apoptosis. Concurrent dysregulation of Hedgehog and Wnt/??-catenin pathways may alter proliferation and survival signals critical for lymphoma cell maintenance. This polyclonal knockout population thus captures heterogeneous functional effects, enabling robust assessment of NEK4 loss in lymphomagenesis and therapeutic resistance.
Researchers can employ these cells for B-cell lymphoma functional studies, including DNA damage response assays (comet assay, ??H2AX foci staining), apoptosis quantification (Annexin V/PI), and cell cycle analysis by flow cytometry. The model is also suited for ciliogenesis research, Hedgehog (GLI reporter) and Wnt (??-catenin/TCF reporter) pathway interrogation, kinase inhibitor screening, and phospho-signaling analysis via phospho-specific antibodies. Co-immunoprecipitation with p53 or drug sensitivity profiling can further elucidate NEK4??s mechanistic roles. For further details, please contact Ascent Research.
