The CYLD Knockout Raji Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout population with targeted disruption of the CYLD gene in the Raji B lymphocyte line. This heterogeneous pool enables loss-of-function studies without clonal selection, reflecting diverse editing outcomes ideal for functional genomics and drug screening in lymphoid contexts.
Derived from an EBV-positive Burkitt lymphoma, the Raji cell line is a hyperdiploid B lymphoblastoid model characterized by constitutively active NF-??B signaling. It retains B-cell receptor signaling features, making it a standard platform for B-cell malignancy research and immune signaling studies. The high basal NF-??B activity sensitizes the system to CYLD-dependent modulation.
CYLD is a K63-specific deubiquitinase that negatively regulates NF-??B, JNK, and Wnt/??-catenin pathways by removing polyubiquitin chains from TRAF2, TRAF6, and RIP1. Upon TNF?? or IL-1?? stimulation, CYLD opposes TAK1-mediated IKK activation, preventing I??B?? phosphorylation and degradation, thereby restraining p65 nuclear translocation. Its tumor-suppressive function involves interactions with NEMO, TAK1, and HDAC6, and it transcriptionally influences targets such as Bcl-3 and c-Myc. Loss of CYLD results in sustained signaling, hyperubiquitination of adaptors, and constitutive pathway activation.
In Raji cells, CYLD knockout exacerbates intrinsic NF-??B and JNK signaling, mirroring oncogenic events in B-cell lymphomas. The enhanced TRAF2/6 ubiquitination and persistent p65 activity in this background highlight the deubiquitinase’s role in preventing unchecked proliferation and survival. This model is thus particularly suited for investigating the interplay between viral oncoproteins and host ubiquitin-editing enzymes during lymphomagenesis.
These polyclonal knockout cells facilitate detailed mechanistic studies using Western blotting of NF-??B effectors (p-I??B??, p65), RNA-seq transcriptomics, flow cytometry for apoptosis and proliferation, and luciferase reporter assays. Co-immunoprecipitation unveils TRAF2/6 ubiquitin status, and phospho-JNK or drug sensitivity screens (e.g., bortezomib) identify CYLD-dependent vulnerabilities. For technical support, contact Ascent Research.
