PEBP1 Knockout Cell Line (HEK293)

The choice depends on whether you are studying PEBP1 (RKIP, Raf kinase inhibitor protein)'s role as a metastasis suppressor or its functions inhibiting Raf-MEK-ERK signaling and IKK/NF-κB pathways. The Knockout line is the standard tool for asking whether RKIP is required for these inhibitory functions — RKIP binds and inhibits Raf-1, MEK1, GRK2, and IKK, broadly attenuating proliferative and inflammatory signaling. Overexpression is useful for studying RKIP tumor suppressor functions in cancer contexts. For cancer biology research, the EDITGENE PEBP1 Knockout in HEK293 enables study of Raf-MEK-ERK and NF-κB pathway hyperactivation following RKIP loss. Rescue with wild-type or phospho-RKIP-mimetic (S153D, which switches RKIP from Raf inhibition to GRK2 inhibition) enables comprehensive structure-function studies. RKIP loss is associated with metastatic progression in multiple cancers — the knockout serves as a tool for studying metastasis-permissive signaling networks.

Primary applications: • Raf-MEK-ERK pathway: phospho-MEK1/2 and phospho-ERK1/2 analysis to assess RKIP-mediated MAPK pathway attenuation in the absence of RKIP. • NF-κB signaling: phospho-IKK, phospho-IκBα, and NF-κB target gene expression analysis given RKIP's IKK inhibition. • GRK2-mediated GPCR desensitization: β-adrenergic receptor desensitization kinetics given RKIP's role in switching from Raf to GRK2 inhibition following S153 phosphorylation. • Metastasis biology: invasion, migration, and EMT marker analysis given RKIP's role as a metastasis suppressor. EDITGENE recommends this model for researchers investigating RKIP-mediated signaling attenuation, metastasis suppression mechanisms, and MAPK/NF-κB pathway regulation.

Yes. RKIP rescue experiments are well-established for signaling research: • Construct design: use a codon-modified PEBP1 sequence with a small C- or N-terminal tag (FLAG, HA). RKIP is small (~21 kDa) cytoplasmic protein — both tag positions are typically tolerated. • Raf-binding-deficient rescue: specific residue mutations in the Raf-binding interface enable separating Raf inhibition from GRK2 inhibition. • Phospho-mimetic rescue: S153D mutation mimics PKC-phosphorylated RKIP, switching from Raf to GRK2 inhibition — invaluable for studying RKIP's dual regulation. • Functional readout: rescue should restore Raf-MEK-ERK pathway attenuation, NF-κB pathway inhibition, and metastasis suppressor phenotypes. HEK293 transduces efficiently with lentivirus and supports stable rescue line generation.