The EEF1A1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting human EEF1A1 in HEK293T cells. This model enables dissection of eukaryotic translation elongation factor 1 alpha 1 functions in translation, cytoskeletal dynamics, and disease processes. The polyclonal pool captures phenotypic diversity from heterogeneous editing, facilitating robust functional studies without single-cell cloning. It is suitable for investigating roles of eEF1A1 in translational control, actin organization, and viral replication.
The HEK293T cell line is a human embryonic kidney derivative optimized for protein expression and viral production. Derived from HEK293 cells via transformation with adenovirus 5 DNA, HEK293T cells express SV40 large T antigen, enhancing episomal replication of SV40 ori?containing plasmids. This permits rapid transient transfection and high-level foreign gene expression, making them ideal for signaling studies, protein interaction analyses, and viral life cycle investigations. Their robust growth and well-characterized proteome ensure a consistent background for loss-of-function experiments.
eEF1A1 serves as a GTP?dependent elongation factor that delivers aminoacyl?tRNAs to the ribosomal A?site. Additionally, it mediates actin bundling, apoptosis regulation, and viral replication via interactions with viral proteins (e.g., HIV?1 Gag, HCV NS5A). mTOR and S6 kinase signaling regulate eEF1A1 activity, integrating nutrient and growth factor cues with protein synthesis. eEF1A1 functions in a complex with eEF1B subunits (EEF1B2, EEF1D, EEF1G) and associates with actin and ribosomes.
Disrupting EEF1A1 in HEK293T cells constructs a model to examine its roles in translation, actin dynamics, and viral production??processes central to the host line??s applications. Effects on global protein synthesis, assessed by puromycin incorporation or polysome profiling, and on actin cytoskeleton organization, visualized by phalloidin staining, can separate eEF1A1??s canonical and non?canonical functions. These insights inform studies on how elongation factor activity influences recombinant protein yield and viral particle assembly.
This polyclonal knockout pool supports a wide range of studies: dissection of mTOR?regulated translation control, co?immunoprecipitation of eEF1A1?associated factors, and investigation of eEF1A1?dependent viral replication. It is compatible with RT?qPCR and Western blotting for expression analysis, cell proliferation assays, and rescue experiments using mutant eEF1A1 constructs. Researchers can map downstream effects on the proteome and screen for pharmacological modulators of elongation factor activity. For further technical details, please contact Ascent Research.
