The EEF1A2 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal loss-of-function population designed to disrupt expression of the EEF1A2 gene in the human renal cell carcinoma line 786-O. This polyclonal knockout model enables functional studies of EEF1A2 in a physiologically relevant cancer background, providing a heterogeneous pool of edited cells for robust experimental analysis.
The host 786-O cell line is derived from a human primary clear cell renal cell adenocarcinoma, representing an epithelial tumor model widely employed in renal cell carcinoma research. Its use allows investigation of oncogenic mechanisms in a cellular context that mirrors key aspects of kidney cancer biology.
EEF1A2 functions as a translation elongation factor that mediates GTP-dependent delivery of aminoacyl-tRNAs to the ribosome, directly coupling protein synthesis to actin cytoskeleton dynamics. Its expression is induced by oncogenic signals including EGFR signaling, c-Myc, and hypoxia via HIF1A, as well as the PI3K/AKT/mTOR pathway. In turn, EEF1A2 promotes translation of cell cycle regulators and anti-apoptotic factors, thereby supporting proliferation and survival. It interacts with ribosomal subunits, actin, aminoacyl-tRNAs, and signaling kinases such as Akt and PI4KIII??, and operates downstream of mTORC1, which phosphorylates S6K and 4E-BP1 to regulate translational control. ERK and AKT cascades further modulate its activity, embedding EEF1A2 within a network that links growth factor signaling to the translational machinery.
In clear cell renal cell carcinoma, EEF1A2 is frequently overexpressed and correlates with enhanced tumorigenic potential. Ablation of EEF1A2 in 786-O cells eliminates the major translation elongation factor isoform, disrupting the synthesis of proteins essential for rapid proliferation and evasion of apoptosis. This loss-of-function model provides a platform to dissect EEF1A2-dependent oncogenic pathways, including mTOR-driven translation and actin-mediated cytoskeletal remodeling, which are critical for tumor cell growth, invasion, and metastasis in kidney cancer.
Researchers can employ this polyclonal knockout population in diverse assays such as western blotting and RT-qPCR to confirm target disruption, MTS and colony formation assays to evaluate proliferation, and annexin V staining to quantify apoptosis. Additionally, the cells are suitable for RNA-seq and polysome profiling to globally assess translational changes, co-immunoprecipitation to map protein interaction networks, and drug sensitivity studies with translation inhibitors like cycloheximide or mTOR inhibitors. These applications enable comprehensive characterization of EEF1A2??s role in renal cell carcinoma and offer a valuable tool for preclinical validation of therapeutic strategies aimed at targeting translation elongation. For further technical information, please contact Ascent Research.
