The EEF1E1 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal HeLa cell population with targeted disruption of EEF1E1, generating a loss-of-function model for studying its roles in translation elongation and p53 regulation. This knockout pool offers a heterogeneous genetic background suitable for population-level studies, avoiding clonal artifacts. The product is intended for functional genomics, pathway analysis, and drug screening in a widely used cervical adenocarcinoma model.
The parental HeLa cell line is an HPV18-positive, aneuploid cervical adenocarcinoma epithelial line established in 1951 from Henrietta Lacks. It is a cornerstone of cancer research, characterized by robust proliferation and ease of manipulation. HeLa cells express wild-type p53, but it is targeted for degradation by HPV E6, providing a permissive environment to explore p53-independent functions of EEF1E1.
EEF1E1 encodes a scaffolding subunit of the multisynthetase complex, which delivers aminoacylated tRNAs to the ribosome via EEF1A1 during translation elongation. EEF1E1 directly binds and stabilizes p53, potentiating its transcriptional activation of p21, BAX, and PUMA. Its activity is regulated by MYC, mTORC1, amino acid levels, and ATM kinase. EEF1E1 interacts with multiple aminoacyl-tRNA synthetases (EPRS, MARS, IARS) and translation elongation factors (EEF1G, EEF1D). Knockout in HeLa cells presumably impairs translation elongation efficiency and may further compromise any residual p53 activity, potentially increasing genomic instability.
In HeLa cells, p53 is functionally inactivated by HPV18 E6, making this knockout model particularly valuable for examining EEF1E1??s contributions to translation control and multisynthetase complex organization independent of p53. This enables the dissection of how compromised translation elongation affects cancer cell growth, survival under genotoxic stress, and drug sensitivity. Researchers can thus study synthetic lethal relationships and the impact on protein homeostasis in a high-turnover cancer context.
Applications include polysome profiling and puromycin incorporation to measure translation rates, Western blotting and immunofluorescence for p21 and apoptotic markers, and flow cytometry for cell cycle and viability assessments. Co-immunoprecipitation and mass spectrometry permit analysis of multisynthetase complex composition. DNA damage response can be evaluated by clonogenic assays, while RNA-seq and ribosome profiling clarify transcriptomic and translatomic alterations. High-throughput screening against protein synthesis inhibitors is also enabled. For technical support, contact Ascent Research.
