EIF5A2 Knockout A2780 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human A2780 ovarian adenocarcinoma cell line, featuring targeted disruption of the EIF5A2 gene. The polyclonal format captures a heterogeneous pool of editing events, avoiding clonal selection bias and providing a representative loss-of-function model for bulk cancer cell analysis. This tool enables interrogation of EIF5A2-dependent phenotypes in a cancer-relevant context.
The A2780 cell line was established from an untreated ovarian carcinoma patient and exhibits epithelial morphology. It retains aberrant mTOR and MYC signaling typical of high-grade serous ovarian cancer, making it a well-suited system for studying oncogenic translation regulation. This line is extensively applied in research on proliferation, adhesion, invasion, and metastasis, offering a relevant background for dissecting EIF5A2 function.
EIF5A2 is a translation elongation factor that resolves ribosome pausing at polyproline motifs, enabling synthesis of proline-rich proteins including Cyclin D1, BCL2, and MMP9. The factor undergoes a unique hypusination modification, catalyzed sequentially by deoxyhypusine synthase (DHPS) and deoxyhypusine hydroxylase (DOHH), which is essential for its activity and subsequent ribosome binding. Upstream, EIF5A2 transcription is activated by MYC, and mTORC1 signaling promotes functional activation through S6K and 4E-BP1. This places EIF5A2 at the intersection of nutrient-sensing and oncogenic translation, where it drives production of proteins critical for tumor progression.
In A2780 cells, EIF5A2 overexpression enhances cell proliferation and metastatic capacity. CRISPR/Cas9-mediated disruption of EIF5A2 abrogates translation of proline-rich oncoproteins, impairing mTOR- and MYC-driven tumorigenic programs. This polyclonal knockout population permits robust assessment of these phenotypes without clonal artifacts, revealing the dependency of ovarian cancer cells on efficient polyproline synthesis. The resulting attenuation of downstream effectors such as Cyclin D1 and BCL2 provides a clear readout for functional studies.
This knockout model supports diverse applications in cancer biology, including translation control studies, functional genomics, and metastasis research. Essential validation assays such as Western blot and RT-qPCR confirm EIF5A2 disruption and downstream target modulation. Functional analyses employ MTT/BrdU proliferation assays, Transwell migration/invasion assays, Annexin V/PI apoptosis detection, colony formation, and polysome profiling to monitor translation efficiency. These methodologies enable comprehensive dissection of EIF5A2-dependent processes. For further technical details or to request a quote, please contact Ascent Research.
