The DPH5 Knockout HGC-27 Polyclonal Cells are a polyclonal knockout cell population generated by CRISPR/Cas9-mediated gene disruption of the DPH5 locus in the HGC-27 human gastric adenocarcinoma cell line. This product provides a heterogeneous pool of edited cells with loss-of-function of DPH5, suitable for studying diphthamide biosynthesis and translation control without clonal selection.
HGC-27 is a poorly differentiated gastric adenocarcinoma cell line originally derived from a metastatic lymph node of a human gastric cancer patient. It serves as a well-established in vitro model for investigating the molecular mechanisms of gastric cancer, including cell proliferation, migration, and response to therapeutic agents. The HGC-27 background is particularly relevant for translational oncology research, enabling the exploration of tumor-specific dependencies on protein synthesis fidelity.
DPH5 encodes a methyltransferase essential for the biosynthesis of diphthamide, a unique post-translational modification of histidine-715 in eukaryotic elongation factor 2 (eEF2). DPH5 catalyzes the trimethylation of the diphthine intermediate, using S-adenosyl methionine as a methyl donor, and functions within a multimeric complex that includes DPH1, DPH2, DPH3, and DPH4. This modification is critical for translational elongation fidelity by preventing ribosomal frameshifting. The activity of DPH5 may be influenced by upstream regulators such as MYC and mTOR signaling, which globally control translation, and its primary downstream target is eEF2, whose diphthamide modification is required for efficient protein synthesis.
In the context of gastric adenocarcinoma, the DPH5 knockout HGC-27 model enables investigation into how diphthamide deficiency impacts cancer cell growth and survival. Given the high proliferative rates of tumor cells and their heightened reliance on protein synthesis, disruption of DPH5 may reveal vulnerabilities that can be exploited therapeutically. Moreover, because diphthamide is the target of ADP-ribosylating toxins such as diphtheria toxin and Pseudomonas exotoxin A, this knockout cell population offers a valuable system to dissect toxin entry, cytotoxicity, and resistance mechanisms in a cancer-relevant background.
This polyclonal knockout cell population is ideal for a range of functional studies, including assessing diphthamide modification status via western blotting, measuring protein synthesis rates with puromycin incorporation, and evaluating susceptibility to diphtheria toxin or Pseudomonas exotoxin A in cell viability assays. Further applications include RT-qPCR or Sanger sequencing to confirm DPH5 disruption, eEF2 immunoprecipitation to probe complex formation, and mass spectrometry for diphthamide detection. The model can also be integrated into functional genomics screens or used to explore the role of translational control in gastric cancer progression. For additional technical details, please contact Ascent Research.
