The DCAF15 Knockout Raji Polyclonal Cells product supplies a CRISPR/Cas9-edited polyclonal population derived from the Raji human B-lymphocyte line, featuring targeted disruption of the DCAF15 gene. This pooled knockout resource circumvents clonal selection, thereby preserving population-level heterogeneity and ensuring representative loss-of-function phenotypes. It is designed for robust and reproducible studies of DCAF15-mediated processes in a biologically relevant lymphoid model.
The Raji host cell line is an Epstein?CBarr virus (EBV)-positive Burkitt lymphoma line of B-cell origin, widely utilized for its stable growth and its ability to model critical B-lymphocyte functions, including antibody secretion, antigen presentation, and immune response. Its transformed state makes it an ideal platform for investigating oncogenic mechanisms and the deregulation of protein homeostasis pathways commonly observed in hematological cancers.
Mechanistically, DCAF15 functions as a substrate-recognition subunit of the CUL4-DDB1-RBX1 E3 ubiquitin ligase (CRL4) complex. It interacts via its WD40 domain with the adaptor protein DDB1, which bridges to the cullin scaffold CUL4A or CUL4B, while RBX1 catalyzes ubiquitin transfer. DCAF15 mediates the polyubiquitination and subsequent proteasomal degradation of target proteins, most prominently the spliceosomal factor RBM39. Its expression is regulated by the transcription factor NRF2. Importantly, the small-molecule aryl sulfonamides indisulam and E7820 co-opt DCAF15 to induce the degradation of neo-substrates such as RBM39, leading to cancer cell apoptosis. Thus, DCAF15 is a critical node linking ubiquitin-dependent proteolysis to pre-mRNA splicing control.
Within the Raji B-lymphoma context, DCAF15 knockout serves as a powerful tool to examine its role in malignant proliferation and survival. Loss of DCAF15 abolishes indisulam-triggered RBM39 degradation, rendering cells resistant to the drug, thereby enabling precise dissection of on-target pharmacological effects. The EBV-positive background further permits exploration of viral?Chost interactions that may modulate CRL4-DCAF15 activity, potentially uncovering dependencies that could be exploited for lymphoma therapy.
This knockout model supports diverse experimental approaches: Western blot analysis of RBM39 degradation, co-immunoprecipitation of DCAF15-DDB1 complexes, cell viability assays with indisulam/E7820, RNA-seq for splicing profiling, and flow cytometry for apoptosis detection. It also enables pooled CRISPR screens to investigate ubiquitin-proteasome regulation. For additional product information, contact Ascent Research.
