This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B-lymphocyte lineage, in which the FRMD8 gene has been disrupted via CRISPR/Cas9-mediated gene editing. The polyclonal knockout format provides a heterogeneous loss-of-function model suitable for studying FRMD8-dependent processes without clonal selection bias. Disruption of the FRMD8 locus yields a mixed population with varied indel mutations that collectively abrogate functional protein expression. This polyclonal approach enables robust assessment of FRMD8 function in a cellular context that more closely mirrors native genetic diversity.
The parental Raji cell line originates from a human Burkitt??s lymphoma and is EBV-positive, maintaining characteristics of mature B lymphocytes, including immunoglobulin production and antigen-presenting capacity. Raji cells grow in suspension with lymphoblastoid morphology, providing a well-established model for B-cell biology, lymphomagenesis, and immune cell signaling. Their EBV-transformed status additionally provides a background relevant to viral-associated lymphoproliferative disorders, where cytoskeletal remodeling and adhesion pathways are often deregulated.
FRMD8 encodes a FERM domain-containing protein predicted to function as a linker between integral membrane proteins and the cortical actin cytoskeleton. FERM domain proteins, such as Ezrin, Radixin, and Moesin (ERM family), mediate membrane-cytoskeleton attachment and are regulated by Rho GTPases and integrin engagement. FRMD8 is proposed to interact with F-actin and plasma membrane components, likely via cytoskeletal adaptors, contributing to actin remodeling downstream of integrin signaling. Although direct upstream regulators remain uncharacterized, the protein may undergo activation through conformational changes upon phospholipid binding or phosphorylation, thereby modulating cell adhesion and migration.
In Raji B lymphocytes, loss of FRMD8 likely impairs dynamic actin reorganization required for adhesion, trafficking, and immune synapse formation. Since Burkitt??s lymphoma cells depend on integrin-mediated interactions for tissue invasion and survival in lymphoid niches, FRMD8 knockout offers a physiologically relevant model to dissect FERM protein contributions to malignant B-cell behavior. Disruption of membrane-to-cytoskeleton linkage may alter adhesion molecule expression and reduce migratory capacity, potentially affecting transendothelial migration and homing to secondary lymphoid organs, key processes in lymphomagenesis and metastasis.
Researchers can use this knockout model in Western blotting and RT-qPCR to confirm FRMD8 ablation and transcript changes, cell adhesion assays to quantify attachment to extracellular matrix or endothelial monolayers, and transwell migration assays to assess chemotactic responses. Immunofluorescence for F-actin and adhesion complex components (e.g., vinculin, paxillin) reveals cytoskeletal defects, while flow cytometry profiles adhesion molecules such as integrins and selectins. Additional applications include co-culture with stromal cells to mimic bone marrow or lymph node environments and drug sensitivity testing to evaluate FRMD8-dependent vulnerabilities. For further information, contact Ascent Research.
