The EEF1D Knockout A-549 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population of A-549 lung epithelial carcinoma cells with targeted disruption of the EEF1D gene. This knockout model system provides a loss-of-function tool for investigating the role of EEF1D in translation regulation, cell proliferation, and oncogenic signaling. The polyclonal format ensures a heterogeneous knockout cell pool, allowing robust functional studies without the clonal selection bias inherent in single-cell-derived lines.
A-549 cells are a well-established model of type II pulmonary epithelium, originally derived from the lung carcinoma tissue of a 58-year-old male. They are widely utilized in lung adenocarcinoma research, epithelial barrier studies, and drug metabolism assays. Their epithelial morphology, adherent growth, and well-characterized signaling networks make them particularly suitable for CRISPR/Cas9-mediated gene disruption studies examining translation factor biology.
EEF1D encodes the delta subunit of the eukaryotic translation elongation factor 1 (eEF1) complex, which functions as a guanine nucleotide exchange factor for EEF1A. By regenerating active EEF1A-GTP, EEF1D facilitates the loading of aminoacyl-tRNAs onto the ribosome during translation elongation. EEF1D activity is regulated by upstream signals including MYC transcription factor, epidermal growth factor (EGF), mTOR kinase, and stress stimuli such as heat shock and oxidative stress. It interacts directly with EEF1A, EEF1B2, EEF1G, and the lysyl-tRNA synthetase KARS, and is modulated by protein kinase C (PKC) and casein kinase 2 (CK2). Downstream, EEF1D promotes global protein synthesis, influencing levels of cyclin D1 and c-Myc, and connects to the mTOR/S6K1 and MAPK/ERK pathways.
Disruption of EEF1D in A-549 cells impairs guanine nucleotide exchange on EEF1A, thereby reducing the pool of active elongation competent ribosomes and attenuating overall protein synthesis. In the lung carcinoma context, this deficiency can compromise cell proliferation, survival under stress, and potentially oncogenic signaling driven by mTOR and MAPK/ERK networks. The resulting polyclonal knockout population provides a physiologically relevant platform to dissect how EEF1D-dependent translation elongation contributes to lung adenocarcinoma pathogenesis and to evaluate the therapeutic potential of targeting the translation machinery.
Typical applications include quantitative analysis of translation rates via puromycin incorporation, phospho-S6 western blotting, and global proteomic profiling. The cells are also suitable for functional assays such as MTT proliferation, colony formation, apoptosis testing, and migration/invasion studies, as well as for co-immunoprecipitation of eEF1 complex components or RNA-sequencing to assess transcriptome-wide changes. This product is thus a valuable resource for cancer researchers investigating translation elongation factors as therapeutic targets. For further details or to place an order, please contact Ascent Research.
