The NEDD4 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Raji B lymphocyte line. This product offers a genetically disrupted NEDD4 gene, enabling loss-of-function studies in a Burkitt??s lymphoma-derived background. The polyclonal nature provides a heterogeneous pool of edited cells, suitable for examining NEDD4-dependent processes without clonal selection artifacts.
Raji cells are an Epstein-Barr virus (EBV)-positive immortalized B cell line originally isolated from a Burkitt??s lymphoma patient. These suspension-adapted cells exhibit a mature B-cell phenotype and are widely employed in immunological and oncology research. The NEDD4 knockout in this context allows dissection of ubiquitin-mediated regulation in B-cell malignancy.
NEDD4 encodes a HECT-type E3 ubiquitin ligase that catalyzes ubiquitination of substrate proteins, primarily targeting them for proteasomal degradation or altered trafficking. NEDD4 regulates key signaling nodes, including PTEN, which controls PI3K/AKT pathway activation, and growth factor receptors such as EGFR and FGFR. Upstream activation involves EGFR and Src family kinases, while NEDD4 activity is modulated by adaptors like NDFIP1 and interaction with the ESCRT machinery. Through ubiquitination of PTEN, NEDD4 downregulates its tumor-suppressive functions, enhancing AKT signaling. Additionally, NEDD4 influences TGF-?? signaling by targeting SMAD proteins, thereby affecting cell proliferation and differentiation.
Within the Raji B-cell lymphoma model, NEDD4 disruption attenuates ubiquitin-dependent regulation of PTEN, likely restoring its lipid phosphatase activity and dampening PI3K/AKT-driven survival signals. This loss-of-function context is instrumental for probing the contribution of NEDD4 to lymphomagenesis, as aberrant ubiquitination events are frequently observed in B-cell malignancies. The polyclonal knockout population enables assessment of heterogeneous editing outcomes on cellular phenotypes such as proliferation and drug sensitivity, providing a more robust representation of genetic perturbation compared to single-cell clones.
Researchers can employ these cells in ubiquitination assays, Western blotting, and co-immunoprecipitation analyses to examine NEDD4 substrate interactions. Functional studies may include proliferation assays, apoptosis detection by flow cytometry, and signaling pathway analysis via phospho-AKT and SMAD phosphorylation readouts. The model also supports drug resistance investigations, particularly related to EGFR or FGFR inhibitors, and is suited for high-throughput screens targeting the ubiquitin-proteasome system in lymphoma. For additional details or custom inquiries, please contact Ascent Research.
