The N4BP1 Knockout Raji Polyclonal Cells product comprises a population of Raji B lymphocytes that have undergone CRISPR/Cas9-mediated disruption of the N4BP1 gene, resulting in a heterogeneous polyclonal knockout cell pool. As a polyclonal population, these cells collectively harbor diverse gene-editing events, providing a robust model for studying population-level effects of N4BP1 loss without the clonal biases present in single-cell-derived lines. The gene disruption abolishes N4BP1 function, establishing a powerful loss-of-function platform for interrogating its cellular roles.
Raji cells are a human B lymphocyte cell line derived from a Burkitt lymphoma patient and retain key characteristics of mature B cells, including surface immunoglobulin expression and antigen-presenting capacity. Their origin in a lymphomagenic context and their inherent NF-??B pathway alterations make them particularly relevant for studying B-cell signaling and immune responses. This host line provides a biologically pertinent background for examining N4BP1??s activities in a B-cell environment.
N4BP1 functions as a critical negative regulator of the NF-??B signaling pathway by interacting with and inhibiting the IKK complex, which consists of IKK??, IKK??, and NEMO. This inhibition blocks I??B?? phosphorylation, thereby retaining the p65/p50 NF-??B dimer in the cytoplasm and dampening transcription of pro-inflammatory target genes such as IL-6 and TNF-??. Upstream, N4BP1 expression is induced by type I interferons (IFN-??/??) and viral infection, positioning it as a feedback attenuator of innate immune responses. Additionally, N4BP1 acts as an interferon-inducible ribonuclease that directly degrades viral RNA, thereby restricting retroviral replication. Its activity is modulated through interactions with E3 ubiquitin ligases NEDD4 and ITCH, which likely regulate its stability or post-translational modifications.
In B lymphocytes, NF-??B signaling is essential for survival, proliferation, and antibody production; its dysregulation is implicated in lymphomagenesis and immune disorders. N4BP1 knockout in Raji cells provides a unique model to dissect how loss of this negative regulator influences NF-??B activation dynamics, cytokine profiles, and cellular responses to viral challenges. Since Burkitt lymphoma is etiologically linked to Epstein-Barr virus infection, this model enables exploration of N4BP1??s antiviral functions within a B-cell context where viral RNA sensing and innate defense are particularly relevant.
Researchers can employ these knockout cells to quantitatively measure NF-??B activity using luciferase reporter assays, evaluate IL-6 and TNF-?? production via ELISA, and assess antiviral capacity through viral replication assays. Comparative RNA-seq between wild-type and N4BP1-disrupted Raji cells can illuminate transcriptomic shifts in interferon-stimulated genes and inflammatory mediators, while flow cytometry facilitates profiling of B-cell activation markers. This polyclonal knockout cell population is well-suited for high-throughput screens and pooled functional studies. For further information or to discuss your specific experimental needs, please contact Ascent Research.
