CUEDC2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Raji B lymphoblast cell line, featuring targeted disruption of the CUEDC2 gene. This heterogeneous pool of gene-edited cells enables functional studies of CUEDC2 loss without clonal isolation, maintaining genetic diversity representative of the parental line. The CRISPR/Cas9-mediated gene disruption ablates full-length CUEDC2 protein expression, providing a tool for investigating NF-??B signaling and ubiquitin-dependent regulation.
The Raji host line is an EBV-positive Burkitt lymphoma-derived B lymphoblast that retains B-cell characteristics such as antibody production and adaptive immune functions. Widely used in B-cell malignancy research, Raji cells model constitutive NF-??B activation driven by EBV latent proteins and autocrine cytokine loops. Their robust growth and genetic tractability make them suitable for CRISPR-editing and downstream functional assays.
CUEDC2 suppresses NF-??B signaling by interacting with the IKK complex (IKK??, IKK??, IKK??/NEMO) to promote dephosphorylation, thereby preventing I??B?? (NFKBIA) degradation and retaining RELA/NFKB1 in the cytoplasm. This inhibits transcription of targets like IL6, TNF, and BCL2. CUEDC2 also interfaces with the ubiquitin-proteasome system via CUL1, SKP1, and ubiquitin, influencing cell cycle regulators CCND1 and CDKN1A, and modulates autophagy.
In Raji cells, CUEDC2 knockout removes a critical NF-??B brake, leading to hypersensitized responses to TNF-?? and IL-1??. This recapitulates deregulated NF-??B signaling common in B-cell lymphomas, enabling studies of proliferation, survival, and drug resistance. The polyclonal format captures functional heterogeneity, especially regarding crosstalk between NF-??B, ubiquitin-mediated proteolysis, and autophagy in EBV-positive lymphoblasts.
Applications include western blotting for phospho-IKK, I??B??, and NF-??B; RT-qPCR profiling of target genes; co-immunoprecipitation of CUEDC2-IKK complexes; ubiquitination assays; and flow cytometry for cell cycle and apoptosis. The cells are suited for luciferase reporter assays and viability screens with proteasome inhibitors. This model accelerates research into NF-??B networks, ubiquitin biology, and lymphoma therapy. Contact Ascent Research for further details.
