This product consists of a CRISPR/Cas9-edited polyclonal knockout cell population of the human renal cell carcinoma line 786-O, engineered for disruption of the EIF4A2 gene. The polyclonal nature provides a heterogeneous pool of edited alleles, enabling functional studies without single-cell clonal isolation. Loss of EIF4A2 expression serves as a loss-of-function model to interrogate cap-dependent translation initiation mechanisms. The knockout population is derived from the parental 786-O epithelial tumor cells and is suitable for comparative analyses with wild-type controls.
786-O is a VHL-mutant clear cell renal carcinoma line originally established from a primary tumor. It retains key characteristics of renal cancer epithelial cells, including activation of hypoxia-inducible pathways downstream of VHL loss. The cell line is widely used as a model for clear cell renal cell carcinoma (ccRCC) and for studying mTOR signaling-dependent translational control. Its genetic background provides a physiologically relevant context for examining the role of translation factors in oncogenesis.
EIF4A2 encodes an ATP-dependent DEAD-box RNA helicase that functions as a core component of the eIF4F translation initiation complex. It unwinds secondary structures within 5?? untranslated regions (UTRs) of mRNAs, facilitating ribosomal scanning and cap-dependent translation. EIF4A2 is regulated by upstream signals including mTORC1, MYC, and PI3K/AKT pathway activity, and interacts directly with EIF4G1, EIF4E, EIF4B, and the inhibitory factor PDCD4. It promotes the translation of downstream targets such as MYC, CCND1, and BCL2, linking growth factor signaling to proliferative and survival programs. Its activity is integrated within the mTOR?CeIF4F axis, together with RPS6KB1, 4E-BPs, and other initiation factors.
In 786-O cells, EIF4A2 contributes to the enhanced translation of structured 5?? UTR oncogenic transcripts, supporting the proliferation and survival of ccRCC tumors. The mTOR?CeIF4F pathway is frequently hyperactivated in renal carcinoma, making this knockout model a valuable tool for dissecting EIF4A2-dependent translational reprogramming. Disruption of EIF4A2 in this VHL-mutant background allows researchers to assess how helicase activity influences the expression of eIF4F-sensitive mRNAs and to evaluate its role as a potential therapeutic vulnerability.
Typical research applications include polysome profiling to monitor translation efficiency changes, western blotting and RT-qPCR for target validation, and colony formation or proliferation assays to assess growth phenotypes. The model can be employed in drug sensitivity studies targeting the eIF4F complex or mTOR signaling, as well as in RNA immunoprecipitation experiments to probe specific mRNA interactions. It further supports investigation of nonsense-mediated decay and global translation control in renal cancer. For technical support or additional details, please contact Ascent Research.
