The DPH5 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa human cervical carcinoma cell line, in which the DPH5 gene has been disrupted to create a loss-of-function model. This polyclonal pool preserves genetic heterogeneity while abolishing DPH5 expression, enabling robust investigation of diphthamide biosynthesis and its broader cellular impacts. The knockout was generated using CRISPR/Cas9-mediated gene disruption, providing a reliable tool for studying the distinct roles of DPH5 in protein synthesis and toxin susceptibility without clonal isolation artifacts.
HeLa cells are an immortalized epithelial line isolated from a cervical adenocarcinoma, making them one of the most widely utilized models in cancer biology and drug discovery. Their rapid proliferation, ease of genetic manipulation, and extensive molecular characterization render them particularly suitable for CRISPR-based knockout studies. In this product, the HeLa background offers a clinically relevant context for examining the consequences of DPH5 loss in a tumor-derived epithelial lineage, where translation regulation is often aberrant.
DPH5 encodes a methyltransferase that catalyzes the trimethylation of the diphthine precursor on histidine 715 of eukaryotic elongation factor 2 (EEF2) to form diphthamide. This conserved post-translational modification is essential for the fidelity of protein synthesis during the elongation phase. DPH5 functions within the diphthamide biosynthetic pathway, which includes DPH1, DPH2, DPH3, DPH4, DPH6, and DPH7, and depends on S-adenosylmethionine as a methyl donor. The diphthamide residue serves as the specific target for ADP-ribosylation by diphtheria toxin and related bacterial ADP-ribosylating toxins, linking DPH5 activity to host cell susceptibility to these pathogenic factors.
In the HeLa cervical carcinoma environment, DPH5 disruption potentially impairs diphthamide synthesis, which may reduce sensitivity to bacterial toxins and alter translational dynamics. Given that dysregulated protein synthesis is a hallmark of cancer, this model enables the dissection of how diphthamide modification contributes to malignant phenotypes and the cellular stress response. HeLa cells expressing oncogenic HPV E6/E7 proteins further allow exploration of crosstalk between viral transformation and translation elongation control, potentially revealing node-specific vulnerabilities in cervical cancer.
Research applications include measuring DPH5 and EEF2 diphthamide levels via western blotting, assessing diphtheria toxin sensitivity to confirm functional knockout, and performing mass spectrometry to quantify diphthamide modification. RT-qPCR can verify DPH5 transcript reduction, while cell proliferation assays evaluate growth effects. CRISPR-mediated rescue experiments using wild-type DPH5 complementation provide rigorous validation. This polyclonal knockout population thus supports diverse studies ranging from basic translation biology to discovery of novel cancer therapeutic targets. For technical specifications or ordering, please contact Ascent Research.
