The EIF1B Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of HeLa cells with targeted disruption of the EIF1B gene, a key translation initiation factor. This heterogeneous pool integrates multiple independent editing events, providing a loss-of-function model free from clonal artifacts and suitable for robust functional studies of translation fidelity in a cancer-relevant context.
The HeLa cell line originated from a cervical adenocarcinoma of Henrietta Lacks and harbors integrated HPV18 sequences. Expressing viral oncoproteins E6 and E7 that inactivate tumor suppressors p53 and Rb, these aneuploid epithelial cells exhibit dysregulated proliferation and elevated protein synthesis rates. As a longstanding model for cancer biology and virology, HeLa offers a well-characterized platform for manipulating translation components to study oncogenic host?Cvirus interactions and stress signaling.
EIF1B functions within the 43S pre-initiation complex as a homolog of eIF1, cooperating with eIF1, the eIF3 complex, eIF5, and the 40S ribosomal subunit to enforce fidelity of AUG start codon recognition. It operates downstream of mTORC1 and the eIF2?? kinases GCN2, PERK, and PKR, which link nutrient availability, growth factor signaling, and cellular stress to translation initiation. Disruption of EIF1B compromises scanning accuracy and alters the translation of transcripts containing upstream open reading frames (uORFs), leading to global proteome shifts and impaired stress-induced translational reprogramming.
Given the HPV-driven dysregulation of mTOR signaling in HeLa cells, EIF1B knockout provides a physiologically relevant model to examine how start-site fidelity impacts oncogenic translation. Loss of EIF1B may exacerbate proteotoxic stress or shift the balance of viral and cellular protein expression, making this tool valuable for dissecting the translation initiation machinery??s role in HPV-mediated carcinogenesis and the integrated stress response. It further enables studies of synthetic lethal interactions and vulnerabilities in cancer cells reliant on high translational output.
Researchers can employ these polyclonal cells for polysome profiling, puromycin incorporation assays, and SILAC proteomics to quantify translation rates and ribosome occupancy. Complementary RNA-seq and RT-qPCR enable transcript-specific analysis of uORF-mediated control, while MTT or CellTiter-Glo assays facilitate drug sensitivity screening against mTOR inhibitors or eIF4F antagonists. For technical specifications or ordering inquiries, please contact Ascent Research.
