The AMFR Knockout A-549 Polyclonal Cells are a polyclonal population of human A-549 lung adenocarcinoma cells engineered via CRISPR/Cas9-mediated gene disruption to ablate expression of the AMFR gene, encoding the gp78 E3 ubiquitin ligase. This knockout model provides researchers with a loss-of-function tool for investigating AMFR-dependent processes in a cancer-relevant context.
The parental A-549 cell line, derived from a 58-year-old Caucasian male with lung carcinoma, is a widely used adherent epithelial model of non-small cell lung adenocarcinoma. These cells harbor an endogenous KRAS G12S mutation and express wild-type p53, recapitulating key oncogenic drivers observed in human lung tumors. The A-549 background offers a clinically relevant platform for studying tumor cell migration, invasion, and drug responses.
AMFR (autocrine motility factor receptor), also known as gp78, is an ER-resident E3 ubiquitin ligase central to ER-associated degradation (ERAD), where it targets misfolded proteins for proteasomal clearance in concert with cofactors including Derlin-1, VIMP, and p97/VCP. Beyond its role in proteostasis, AMFR serves as a receptor for autocrine motility factor (AMF/PGI); ligand binding activates PI3K/AKT signaling, leading to phosphorylation of downstream effectors such as mTOR, S6K, and the Rho GTPases Rac1 and Cdc42, which regulate actin dynamics and cell motility. AMF/AMFR signaling also upregulates matrix metalloproteinases MMP-2 and MMP-9 and induces transcription factors such as Snail, thereby driving epithelial-mesenchymal transition (EMT).
In the A-549 lung adenocarcinoma context, AMFR knockout disrupts both ERAD function and AMF-mediated pro-metastatic signaling. A-549 cells express AMFR and AMF, and autocrine/paracrine AMF/AMFR signaling contributes to their migratory and invasive phenotype. Ablation of AMFR in this polyclonal population thus attenuates key metastatic behaviors, making the model valuable for dissecting how ERAD and AMF/AMFR pathways intersect with oncogenic KRAS signaling to promote tumor progression.
This knockout cell pool is suitable for a wide range of experimental applications, including wound healing and transwell migration/invasion assays to assess motility, western blotting for AMFR, AKT, and phospho-AKT to probe signaling, co-immunoprecipitation of ubiquitinated substrates to evaluate ERAD activity, and fluorescence-based proteasome activity measurements. The model further supports chemotaxis studies, small-molecule inhibitor screening targeting gp78, and EMT marker expression analysis. Researchers may employ these cells to validate AMFR as a therapeutic target in KRAS-mutant lung adenocarcinoma or to explore crosstalk between ubiquitin-proteasome deregulation and oncogenic signaling. For additional information or technical support, please contact Ascent Research.
