The EEF1E1 Knockout HT29 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population of HT29 cells with targeted disruption of the EEF1E1 gene. This heterogeneous knockout model avoids clonal artifacts and is ideal for studying EEF1E1-dependent functions in a mixed genetic background.
The HT29 cell line is a human colorectal adenocarcinoma model established from a 44-year-old female. It exhibits adherent epithelial morphology, KRAS wild-type status, mutant p53, and the capacity for enterocytic differentiation, making it a standard system for intestinal tumor biology and differentiation research.
EEF1E1 (AIMP3) is a tumor suppressor and component of the multi-tRNA synthetase complex that links translation elongation to cell fate. Upon genotoxic or nutrient stress, EEF1E1 dissociates from the complex, translocates to the nucleus, and activates p53 to induce p21 and PUMA, promoting apoptosis or cell cycle arrest. Concurrently, it stabilizes PTEN to inhibit AKT signaling, thereby reducing cell survival. The protein is regulated by AKT-mediated phosphorylation that triggers degradation, and its levels respond to growth factors like EGF and amino acid availability. EEF1E1 interacts with p53, PTEN, JAB1 (COPS5), and synthetases such as EPRS, KARS, and RARS, integrating translational activity with tumor-suppressive pathways.
In HT29 cells, which carry a p53 mutation, EEF1E1 knockout enables dissection of p53-independent tumor-suppressive mechanisms and crosstalk with PTEN/AKT signaling. Loss of EEF1E1 may relieve inhibition on AKT and reduce apoptotic priming, potentially mimicking oncogenic states in colorectal cancer where EEF1E1 function is compromised. This model is thus particularly relevant for studying how translational control interfaces with growth signaling in colorectal adenocarcinoma.
Research applications include investigation of EEF1E1’s role in colorectal cancer, analysis of p53-independent apoptosis, evaluation of chemosensitivity to drugs like 5-fluorouracil, and study of translation dysregulation in tumorigenesis. Compatible assays range from Western blotting and RT-qPCR for knockout validation, to co-immunoprecipitation with p53 or PTEN, flow cytometric apoptosis detection (Annexin V/PI), polysome profiling, and migration/invasion experiments. For further details, contact Ascent Research.
