The NSFL1C Knockout Raji Polyclonal Cells product provides a heterogeneous population of Raji B lymphoblastoid cells bearing CRISPR/Cas9-mediated disruption of the NSFL1C gene. This polyclonal knockout approach avoids clonal selection artifacts, preserving genetic variability typical of edited pools. The resulting loss-of-function model enables robust investigation of p47-dependent processes without the need for single-cell cloning.
Raji cells originate from a Burkitt lymphoma patient and maintain latent Epstein-Barr virus (EBV) infection, making them a widely adopted system for studying B cell malignancies and viral biology. As a lymphoblastoid line, Raji cells display activated B cell markers and active growth signaling, rendering them suitable for probing protein quality control mechanisms that intersect with oncogenic pathways and immune receptor trafficking.
The NSFL1C gene encodes p47, a UBX domain-containing cofactor that recruits the AAA+ ATPase p97/VCP to ubiquitinated substrates on membranes. p47 orchestrates key processes including endoplasmic reticulum-associated degradation (ERAD), autophagosome maturation, and mitotic Golgi reassembly. Mechanistically, p47 forms ternary complexes with VCP and ubiquitin to extract ubiquitinated proteins from membranes, facilitating their proteasomal degradation or membrane fusion events. This activity is regulated by CDK1/cyclin B phosphorylation during mitosis and by ER stress, and p47 directly interacts with Golgi matrix proteins GM130 and GRASP65, syntaxin 5, and competes with the UFD1/NPL4 adaptor complex for VCP binding. Downstream, p47 controls the fate of ERAD substrates, autophagic cargo, and Golgi reassembly intermediates, thus integrating the ubiquitin-proteasome system with membrane dynamics.
Within the Raji lymphoma environment, NSFL1C knockout disrupts p47-dependent proteostasis pathways that are often essential for malignant B cell survival. This model allows dissection of how loss of this cofactor alters ERAD efficiency, autophagy flux, and Golgi morphology, potentially sensitizing cells to proteotoxic stress or proteasome inhibitor therapy. It is particularly valuable for exploring p97/cofactor dependencies in hematological cancers and for elucidating connections between protein quality control and EBV-driven oncogenesis.
Research applications span a range of biochemical and cell-based assays: western blotting to monitor p47, p97, and ubiquitinated protein levels; co-immunoprecipitation to examine p97?Ccofactor interactions; flow cytometry for apoptosis and B cell surface markers; immunofluorescence microscopy to visualize Golgi fragmentation and autophagosome accumulation; and drug sensitivity profiling with proteasome inhibitors such as bortezomib. These polyclonal knockout cells are also suitable for pooled CRISPR screens and chemical biology studies targeting protein degradation networks in B cell cancers. For technical support or customization requests, please contact Ascent Research.
