The DPH5 Knockout MES-OV Polyclonal Cells provide a heterogeneous population of CRISPR/Cas9-edited MES-OV ovarian cancer cells with targeted disruption of the DPH5 gene. This polyclonal format preserves diverse editing outcomes, enabling studies that account for variable knockout efficiencies and potential off-target effects within a biologically relevant cancer cell background.
The host MES-OV cell line is an epithelial ovarian cancer model derived from a metastatic lesion of a mesonephric-like adenocarcinoma, a rare and aggressive subtype. These cells harbor a TP53 mutation, mirroring a prevalent genetic alteration in high-grade serous ovarian carcinomas. The mesonephric-like phenotype is associated with unique transcriptional programs and signaling dependencies, making MES-OV a valuable platform for preclinical research on ovarian cancer pathogenesis and drug response.
DPH5 encodes a methyltransferase that catalyzes the trimethylation of histidine 715 on eukaryotic elongation factor 2 (eEF2), the penultimate step in diphthamide biosynthesis. This modification is critical for translational fidelity and constitutes the target for ADP-ribosylation by bacterial toxins such as diphtheria toxin and Pseudomonas exotoxin A. DPH5 functions within a multienzyme complex comprising DPH1, DPH2, DPH3, and DPH4, and utilizes S-adenosylmethionine as a methyl donor. Its expression is driven by constitutive and MYC-regulated transcriptional programs, linking diphthamide synthesis to general translation control. Downstream, diphthamide-modified eEF2 influences translation elongation dynamics, and its ADP-ribosylation by toxins results in irreversible inhibition of protein synthesis and cell death.
In the TP53-mutant MES-OV background, loss of DPH5 may disrupt translation elongation fidelity, potentially affecting oncogenic protein expression and stress adaptation. This model allows dissection of interactions between translation control pathways and p53-mediated tumor suppression. Because diphthamide is the receptor for ADP-ribosylating toxins, DPH5 knockout confers resistance to toxin-induced killing, enabling studies of toxin-based therapeutic strategies and identification of synthetic vulnerabilities in ovarian cancer. The polyclonal population better mimics intercellular heterogeneity observed in tumors compared to clonal lines.
Researchers can utilize these cells in diverse assays: western blotting with anti-eEF2 and anti-diphthamide antibodies to confirm loss of modification; dose-response experiments with diphtheria toxin or Pseudomonas exotoxin A to quantify functional knockout; polysome profiling or puromycin incorporation to measure translation elongation rates; RNA-seq or ribosome profiling to assess global translational changes; immunofluorescence microscopy to examine eEF2 localization; and MTT or other viability assays to evaluate growth phenotypes under various conditions. These applications support investigations into ovarian cancer biology, protein synthesis regulation, and susceptibility to microbial toxins. For additional technical specifications or ordering information, please contact Ascent Research.
