OTUD6B Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the OTUD6B gene has been disrupted in Raji B lymphocytes. This heterogeneous pool of cells provides a loss-of-function model for studying the deubiquitinase OTUD6B, circumventing clonal selection biases. The CRISPR/Cas9-mediated gene disruption enables robust interrogation of OTUD6B-dependent pathways in a physiologically relevant B-cell context.
Raji cells are human Burkitt’s lymphoma-derived B lymphocytes that are Epstein-Barr virus (EBV)-positive and exhibit constitutive NF-??B activity, making them a well-established model for B-cell lymphoma and antigen presentation studies. The oncogenic and viral milieu creates a sensitized background where negative regulators of NF-??B, such as OTUD6B, can be functionally interrogated.
OTUD6B encodes a deubiquitinase that specifically removes K63-linked polyubiquitin chains from substrates like TRAF3, a pivotal adaptor in innate immune signaling. Under stimulation by upstream regulators TNF-??, IL-1??, or TLR ligands, TRAF3 ubiquitination orchestrates activation of the IKK complex (IKK??/IKK??/IKK??), leading to I??B?? phosphorylation, degradation, and NF-??B (p65) nuclear translocation. OTUD6B deubiquitination of TRAF3 counteracts this cascade, attenuating transcription of pro-inflammatory cytokines such as IL-6 and TNF-??. The enzyme also interacts with TRAF6 and ubiquitin ligases cIAP1/2, thereby fine-tuning ubiquitin-mediated signal termination. Consequently, OTUD6B disruption unleashes NF-??B signaling, promoting cytokine expression and cell survival.
In Raji B-lymphoma cells, OTUD6B loss amplifies the already elevated NF-??B signaling driven by EBV and oncogenic lesions, mimicking lymphomagenic conditions of chronic inflammation and apoptotic resistance. This polyclonal model captures diverse knockout-induced phenotypes, enabling robust correlation of deubiquitinase activity with malignant B-cell traits. Moreover, the model extends to studies of intellectual disability syndrome and microcephaly by providing an immune cell platform to examine OTUD6B??s broader roles in NF-??B-dependent developmental processes.
These knockout cells are applicable to researching OTUD6B??s role in B-cell lymphoma, NF-??B pathway regulation, antiviral innate immunity, and inflammatory gene expression. Representative techniques include Western blotting of OTUD6B, TRAF3, I??B??, and p65; RT-qPCR analysis of IL-6, TNF-??, and IFN-??; NF-??B luciferase reporter assays; co-immunoprecipitation of ubiquitinated TRAF3; flow cytometry for apoptosis and proliferation; and ELISA for cytokine secretion. For additional details, please contact Ascent Research.
