The EIF4EBP1 Knockout Ca Ski Polyclonal Cells are a CRISPR/Cas9-mediated polyclonal knockout cell population targeting the EIF4EBP1 gene in the human cervical carcinoma Ca Ski cell line. This gene-edited pool enables loss-of-function studies of the translational repressor 4E-BP1 without clonal selection, providing a heterogeneous population that reflects varied editing outcomes for robust functional analyses.
The Ca Ski cell line, derived from a cervical epidermoid carcinoma metastasis, serves as an epithelial model of HPV16-positive cervical cancer. These adherent cells express HPV16 E6 and E7 oncoproteins, which inactivate p53 and pRb, driving uncontrolled proliferation. The genomic integration of HPV16 and the epithelial morphology make Ca Ski particularly relevant for translational research in HPV-driven malignancies.
EIF4EBP1 encodes 4E-BP1, a key translational repressor that binds eIF4E to inhibit cap-dependent translation initiation. This interaction is regulated by mTORC1 phosphorylation in response to upstream signals from the PI3K-AKT pathway, activated by insulin/IGF1 receptors (INSR, IGF1R) and transduced through RHEB and TSC1/TSC2. Dephosphorylated 4E-BP1 sequesters eIF4E, blocking assembly of the eIF4F complex; upon mTORC1-mediated phosphorylation, it releases eIF4E, allowing translation of targets such as cyclin D1, c-Myc, VEGF, Mcl-1, and survivin. mTORC1 components including raptor, mLST8, and DEPTOR interact dynamically with 4E-BP1, positioning it as a central effector of growth-promoting translation.
In HPV16-positive Ca Ski cells, disruption of 4E-BP1 via CRISPR/Cas9 likely amplifies eIF4E-dependent translation, synergizing with HPV oncoprotein-mediated activation of the PI3K-AKT-mTOR pathway. This knockout model is therefore instrumental for studying the reliance of cervical cancer cells on mTORC1-driven translation for proliferation and survival, and for investigating mechanisms of resistance to mTOR-targeted therapies. The polyclonal knockout population also reflects tumor heterogeneity, enabling the study of variable translational responses within a cancer cell mass.
Researchers can utilize these cells for western blot analysis of 4E-BP1 and phospho-4E-BP1, cap-binding assays, and polysome profiling to assess translation regulation. Functional assays include proliferation, colony formation, migration/invasion, and apoptosis flow cytometry. The model supports mTOR inhibitor sensitivity testing and exploration of HPV oncoprotein effects on translation. It also aids biomarker discovery for cervical cancer prognosis. For additional details, please contact Ascent Research.
