The EIF5A2 Knockout 786-O Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population derived from the 786-O human renal adenocarcinoma cell line, designed to disrupt the EIF5A2 gene and generate a heterogeneous loss-of-function model. This polyclonal knockout cell pool preserves the genetic variation inherent to CRISPR-based gene disruption, facilitating robust functional analyses while avoiding clonal selection biases.
The 786-O cell line is an established human kidney epithelial model originating from a primary clear cell renal cell carcinoma. Characterized by constitutive activation of mTOR signaling and MYC transcriptional networks, 786-O cells serve as a standard platform for investigating renal cell carcinoma biology, particularly translational dysregulation and tumor cell proliferation.
EIF5A2 encodes a translation elongation factor that promotes peptide bond formation, enhancing synthesis of proteins involved in cell cycle progression (cyclin D1) and apoptosis inhibition (Bcl-2). Its expression is regulated by MYC and HIF1A, functioning downstream of mTORC1 alongside S6K and 4E-BP1. EIF5A2 interacts with ribosome subunits and deoxyhypusine synthase, and also influences MMP9 expression, thereby contributing to invasive phenotypes. Disruption of EIF5A2 uncouples translation from oncogenic signaling, reducing pro-survival protein output. Consequently, EIF5A2 knockout leads to attenuated synthesis of proteins encoded by MYC target mRNAs and diminished translational output downstream of mTORC1.
In 786-O cells, EIF5A2 knockout attenuates the translation of mTOR/MYC-dependent mRNAs, leading to diminished proliferation, increased apoptosis, and impaired colony formation. This model captures the dependency of renal carcinoma on elongation factor-driven protein synthesis and provides a physiologically relevant system to study how HIF1A and growth factor pathways converge on the translation machinery. Moreover, investigators can use this knockout model in combination with mTOR pathway inhibitors to explore synergistic anti-proliferative effects.
This polyclonal knockout cell population is suited for a range of assays, including western blotting for EIF5A2 and downstream targets, MTT-based proliferation assays, Annexin V apoptosis detection, RT-qPCR, colony formation assays, and xenograft tumor studies. Transcriptomic analyses via RNA-seq and polysome profiling can capture global translation effects. Key applications include functional validation of EIF5A2 as an oncogene, interrogation of mTOR/MYC-driven translational control, and screening for small-molecule inhibitors. For further information, please contact Ascent Research.
