CUL4A Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population with disrupted CUL4A gene expression in the Raji B lymphoblastoid line. This pool of edited cells provides a robust loss-of-function model without clonal isolation, suitable for bulk biochemical and functional assays.
The Raji host cell line is an Epstein-Barr virus (EBV)-positive B lymphoblastoid line derived from a Burkitt lymphoma patient. These cells are extensively used in immunology and oncology research because of their robust in vitro growth, expression of B-cell markers, and inherent EBV-driven signaling, making them a relevant model for studying B-cell malignancies, viral oncogenesis, and immune cell biology.
CUL4A serves as a molecular scaffold for the CUL4A-RING E3 ubiquitin ligase (CRL4A) complex. It interacts with the adaptor protein DDB1 and various DCAF substrate receptors to recruit specific targets, including CDT1 (a DNA replication licensing factor), the CDK inhibitor p21, the transcription factor HOXA9, and the DNA repair factor DDB2. The RING finger protein RBX1 catalyzes ubiquitin transfer, marking substrates for proteasomal degradation. CRL4A activity is tightly regulated: NEDD8 conjugation by the NAE1/UBA3 enzyme activates the complex, while the COP9 signalosome mediates deneddylation, and CAND1 facilitates dynamic cullin exchange. Upstream DNA damage signals from ATM/ATR kinases and the tumor suppressor p53 modulate CUL4A function, linking it to cell cycle checkpoints and genome stability.
In the Raji B-cell context, CUL4A knockout enables detailed investigation of its contributions to lymphomagenesis. Dysregulated CRL4A activity has been associated with Burkitt lymphoma, breast, lung, and hepatocellular carcinomas. This polyclonal knockout model allows researchers to examine how loss of CUL4A affects ubiquitin-mediated proteolysis, cell cycle control, and DNA repair in an EBV-positive environment. It is particularly valuable for studying mechanisms of B-cell transformation, viral latency, and resistance to DNA-damaging agents, as well as for evaluating the oncogenic role of CUL4A in hematopoietic malignancies.
Typical experimental approaches with this product include Western blotting to confirm CUL4A depletion and monitor downstream targets like CDT1 and p21; proliferation assays (MTS); cell cycle analysis by propidium iodide staining and flow cytometry; apoptosis detection with annexin V; and DNA damage assessment via ??H2AX immunofluorescence. Interaction studies using co-immunoprecipitation can assess the integrity of the CRL4A complex, and in-vivo ubiquitination experiments validate substrate modification. Transcriptomic analyses with RNA-seq and RT-qPCR enable gene expression profiling. These tools support drug target validation for CRL4A inhibitors, functional dissection of ubiquitin signaling in B-cell malignancies, and host-pathogen interaction studies with EBV. For additional details, please contact Ascent Research.
