The EEF2K Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal human cell population with targeted disruption of the EEF2K gene (eukaryotic elongation factor 2 kinase). This loss-of-function model is generated in the widely used HEK293T background and is provided as a heterogeneous pool of edited cells, enabling researchers to investigate EEF2K-dependent regulation of translational elongation and cellular stress responses without clonal selection artifacts. The polyclonal format supports pooled screening applications and facilitates rapid assessment of downstream molecular phenotypes.
HEK293T cells are human embryonic kidney epithelial cells transformed with adenovirus 5 DNA, which also express the SV40 large T-antigen. These cells exhibit high transfection efficiency, making them a preferred host for ectopic gene expression and CRISPR-based genome engineering. Their renal proximal tubule epithelial origin and robust growth characteristics make them suitable for a broad range of biochemical and cell signaling studies, including those focused on nutrient sensing, growth factor signaling, and autophagy.
EEF2K functions as a calcium/calmodulin-dependent kinase that phosphorylates EEF2 at Thr56, thereby inhibiting ribosomal translocation and reducing protein synthesis elongation rates. This kinase integrates regulatory inputs from mTORC1 and AMPK to coordinate cell growth with nutrient and energy availability. Under conditions of metabolic stress, EEF2K activation promotes autophagy via modulation of ULK1 and LC3-associated autophagic flux, while also contributing to cell cycle arrest. The enzyme is further regulated by cAMP/PKA and MAPK/ERK pathways, linking mitogenic and stress signals to translation control.
In the HEK293T background, disruption of EEF2K provides a powerful tool for dissecting the interplay between protein synthesis and cellular adaptation mechanisms. Given the high transfection efficiency of these cells, researchers can easily complement the knockout with wild-type or mutant EEF2K constructs to validate functional domains. The model is relevant for studying solid tumor biology, where EEF2K supports survival under hypoxic and nutrient-deprived conditions, as well as for neurodegenerative disease research involving dysregulated protein homeostasis. Additionally, the system enables analysis of calcium-dependent signaling circuits that converge on elongation control.
Typical applications include western blotting for phospho-EEF2 (Thr56) to confirm loss of kinase activity, puromycin incorporation assays to measure global protein synthesis, and autophagy flux assays using LC3-II turnover. The cells are also suited for viability assays under metabolic stress conditions, such as glucose or amino acid deprivation, and for co-immunoprecipitation studies examining EEF2K?Ccalmodulin interactions. Drug target validation efforts for inhibitors of EEF2K in cancer or cardiac hypertrophy further benefit from this model. For further product details, please contact Ascent Research.
