The ERRFI1 Knockout Raji Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population in which the ERRFI1 gene has been disrupted, eliminating its regulatory function. This polyclonal pool enables broad assessment of gene disruption effects without clonal selection artifacts, serving as a versatile loss-of-function model for studying negative feedback control of EGFR/ERBB receptor tyrosine kinase signaling.
The host cell line, Raji, is a human B lymphoblast line originally derived from a Burkitt’s lymphoma patient. These suspension cells exhibit antigen-presenting cell properties and are widely employed as a model system in B-cell biology, immunology, and oncology research. Their rapid proliferation and well-characterized signaling landscape make them suitable for investigating oncogenic and immune-related pathways.
ERRFI1 (also known as MIG-6) acts as a critical negative regulator of EGFR/ERBB receptors by directly binding to their kinase domains, inhibiting catalytic activity, and promoting receptor internalization. This interaction suppresses downstream cascades including the RAS-MAPK pathway, where GRB2, SOS, RAS, RAF, MEK1/2, and ERK1/2 are sequentially activated, and the PI3K-AKT pathway involving AKT1/2/3. Additionally, ERRFI1 influences JNK signaling and associates with adaptors such as 14-3-3 proteins and small GTPases CDC42 and RAC1. Knockout of ERRFI1 abolishes this inhibitory feedback, leading to sustained or exaggerated signaling upon stimulation by EGF, TGF-alpha, or other ERBB ligands.
In the Raji B-cell context, loss of ERRFI1-mediated attenuation renders the cells hypersensitive to EGFR/ERBB pathway activation. This can drive altered proliferation, survival, and immune-related functions, potentially impacting cell cycle regulators like p21 and p27 and apoptotic mediators such as BAD and BIM. The model thus provides a valuable tool to dissect how disruption of negative feedback rewires signaling networks unique to B-lymphoid malignancies and modulates responses to external stimuli.
This knockout cell population supports diverse experimental applications, including western blot analysis of phospho-EGFR, phospho-ERK, and phospho-AKT to assess pathway activity, RT-qPCR profiling of ERRFI1 and downstream target genes, cell proliferation and apoptosis assays, flow cytometric measurement of surface EGFR, co-immunoprecipitation to probe EGFR-adaptor interactions, and drug sensitivity screening with EGFR inhibitors. It is suited for research areas such as EGFR/ERBB-driven cancers, non-small cell lung cancer, glioblastoma, and inflammatory diseases. For additional specifications or protocol guidance, please contact Ascent Research.
