The GID8 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Raji human B lymphocyte line, featuring disruption of the GID8 gene to create a loss-of-function model. This heterogeneous pool enables robust functional studies without clonal selection bias, allowing investigation of GID8-dependent pathways in a B-cell malignancy context.
Raji cells are an EBV-positive Burkitt lymphoma line of B-cell origin that grows in suspension with lymphoblastoid morphology. Widely utilized to model humoral immunity and lymphomagenesis, they provide a pathophysiologically relevant system for studying metabolic and proteostatic perturbations in B-cell lymphoma. Their ease of genetic manipulation and rapid proliferation make them ideal for CRISPR-based knockout experiments.
GID8 functions as a substrate recognition subunit of the CTLH E3 ubiquitin ligase complex, which targets proteins such as HMGCS1 for ubiquitination and proteasomal degradation. The CTLH complex, composed of WDR26, GID4, ARMC8, MAEA, RMND5A, and other subunits, integrates nutrient and metabolic stress signals to control substrate turnover. GID8 activity is modulated by upstream metabolic cues and is essential for regulating the mevalonate pathway through HMGCS1 degradation, linking ubiquitin-proteasome activity to cholesterol biosynthesis and metabolic homeostasis.
In Raji B cells, GID8 knockout impairs CTLH-mediated degradation, leading to HMGCS1 accumulation and potential dysregulation of cholesterol metabolism. This may alter membrane dynamics, protein prenylation, and signaling that support lymphoma proliferation. Furthermore, disruption of proteostatic control can affect cell cycle progression, apoptosis, and adaptive responses to metabolic stress, highlighting vulnerabilities in B-cell malignancies that depend on efficient protein turnover. The model thus enables elucidation of how the CTLH complex influences lymphoma cell biology.
These polyclonal knockout cells are suitable for studying GID8-dependent ubiquitination, substrate identification, and metabolic regulation in B lymphocytes. Compatible applications include Western blotting for HMGCS1 and CTLH subunits, ubiquitination assays, RT-qPCR, cell cycle analysis, apoptosis assays, cholesterol quantification, and drug sensitivity profiling. They support investigations into B-cell receptor signaling, synthetic lethal interactions, and high-throughput screening. For further details or to order, contact Ascent Research.
