The FBXO3 Knockout Raji Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population in which the FBXO3 gene has been disrupted in the human Raji B lymphocyte cell line. This polyclonal knockout model is generated via CRISPR/Cas9-mediated gene disruption, yielding a heterogeneous pool of cells carrying loss-of-function mutations at the FBXO3 locus. The resulting knockout population serves as a powerful experimental system to dissect FBXO3-dependent signaling and its roles in B-cell biology and lymphomagenesis without monoclonal selection, thereby preserving phenotypic diversity that may reflect physiological heterogeneity.
The parental Raji cell line is derived from a Burkitt lymphoma patient and harbors the Epstein-Barr virus (EBV) genome. These suspension-adapted B lymphocytes retain antigen-presenting capabilities and are widely employed as a model for EBV-associated B-cell lymphomas. The EBV-positive background drives constitutive activation of survival and proliferation pathways, including NF-??B signaling, making Raji cells particularly suitable for interrogating oncogenic networks that underpin B-cell malignancies.
FBXO3 functions as the substrate recognition subunit of the SKP1-CUL1-RBX1 (SCF) E3 ubiquitin ligase complex. Within this SCF^FBXO3 assembly, FBXO3 selectively recruits the substrate FBXL2 for ubiquitination and subsequent proteasomal degradation. Degradation of FBXL2 stabilizes cyclin D2, which partners with CDK4 and CDK6 to promote G1/S cell cycle progression. Concurrently, FBXO3-mediated turnover of FBXL2 and possibly TRAF2/TRAF6 relieves negative regulation of NF-??B, enabling RELA/p65 transcriptional activity. Thus, FBXO3 operates as a positive regulator of cell cycle entry and NF-??B survival signaling, integrating upstream cues from MYC and NF-??B itself. Interacting factors such as SKP1, CUL1, and RBX1 constitute the core ligase machinery, while downstream targets include the tumor suppressors FBXL2 and the adaptor proteins TRAF2/TRAF6.
In the context of Raji cells, FBXO3 ablation disrupts a critical oncogenic hub that simultaneously drives proliferation and anti-apoptotic programs. Lymphoma cells constitutively deploy SCF^FBXO3 activity to accelerate cyclin D2 accumulation and sustain NF-??B signaling, both hallmarks of Burkitt lymphomagenesis. Therefore, this knockout model allows researchers to investigate the dependence of EBV-transformed B cells on the ubiquitin-proteasome system, to dissect cross-talk between cell cycle machinery and inflammatory signaling, and to evaluate the therapeutic potential of targeting the SCF^FBXO3 complex.
Key applications include western blotting for monitoring cyclin D2, FBXL2, and phospho-p65 levels; co-immunoprecipitation to confirm interactions between FBXO3 and SCF components; ubiquitination assays to track FBXL2 modification; flow-cytometric cell cycle analysis; NF-??B luciferase reporter assays; and proliferation assays under various stimuli. This knockout polyclonal population supports drug target validation studies, cancer biology investigations, and mechanistic research into ubiquitin-proteasome regulation in B-cell malignancies. For further technical information or inquiries about custom solutions, please contact Ascent Research.
