The EIF4EBP1 Knockout 786-O Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout population of 786-O human kidney epithelial cells harboring targeted disruption of the EIF4EBP1 gene. This loss-of-function model stably ablates the 4E-BP1 translational repressor, enabling dissection of cap-dependent translation control in renal cell carcinoma (RCC). The polyclonal nature captures a heterogeneous pool of gene-edited cells, facilitating robust population-level analyses of mTOR pathway signaling and downstream growth phenotypes without clonal selection bias.
The 786-O cell line originates from a human clear cell renal cell carcinoma (ccRCC) and carries a biallelic VHL mutation, resulting in constitutive HIF stabilization and aberrant activation of the PI3K?CAKT?CmTOR signaling axis. This epithelial line retains key renal cell characteristics, making it a highly relevant model for studying the molecular pathology of ccRCC. The VHL-defective background also engenders metabolic reprogramming and heightened sensitivity to mTOR pathway manipulation, providing a disease-appropriate context for EIF4EBP1 knockout studies.
The EIF4EBP1 gene encodes 4E-BP1, a translational repressor that binds eIF4E to inhibit cap-dependent translation initiation. mTORC1, activated by upstream signals including AKT, PI3K, insulin/IGF-1 receptors, and amino acid sensing, phosphorylates 4E-BP1 via interaction with RPTOR. This phosphorylation releases eIF4E, derepressing translation of oncogenic mRNAs such as cyclin D1, c-Myc, VEGF, survivin, Mcl-1, and ODC. 4E-BP1 operates downstream of the TSC1?CTSC2?CRHEB module and integrates signals from the insulin and MAPK pathways to control cell growth.
In the VHL-mutant 786-O model, loss of 4E-BP1 is predicted to enhance cap-dependent translation of growth-promoting mRNAs, potentially exacerbating tumorigenic properties and altering sensitivity to mTOR inhibitors like rapamycin. This knockout system is ideal for delineating the specific roles of 4E-BP1 versus other mTORC1 effectors in ccRCC progression. It also provides a platform to investigate how translational reprogramming contributes to drug resistance, a critical challenge in renal cancer therapy.
These EIF4EBP1 knockout polyclonal cells support a wide range of assays, including western blotting for total and phospho-4E-BP1, eIF4E co-immunoprecipitation, polysome profiling, and RNA-seq. Functional assays include MTT proliferation, colony formation, and rapamycin sensitivity testing. They also facilitate high-throughput screens for mTORC1-dependent synthetic lethal interactions. For detailed product information or to discuss your research needs, please contact Ascent Research.
