The DUS3L Knockout HGC-27 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout population with targeted disruption of the DUS3L gene in the HGC-27 human gastric adenocarcinoma cell line. This polyclonal format provides a heterogeneous mixture of edited cells, enabling robust loss-of-function studies without the biases inherent to monoclonal selection. The model is specifically designed to interrogate the role of tRNA modifications mediated by dihydrouridine synthase in a malignant gastric epithelial context.
HGC-27 is an epithelial cell line established from the metastatic lymph node of a 45-year-old male with gastric adenocarcinoma. These cells retain key characteristics of aggressive gastric carcinoma, including metastatic potential, and are extensively used as an in vitro system for investigating gastric cancer pathogenesis, drug response, and molecular mechanisms underlying tumor progression.
DUS3L functions as a tRNA dihydrouridine synthase that utilizes NADPH to reduce uridine to dihydrouridine at specific positions in the D-loop of tRNA molecules. This post-transcriptional modification is essential for preserving tRNA structural flexibility and thermodynamic stability, which in turn ensures high-fidelity codon?Canticodon interactions during ribosomal translation. DUS3L operates within a network regulated by transcriptional controllers of tRNA synthesis such as MYC and RNA polymerase III, and its catalytic activity directly influences the elongation phase of translation via factors like eEF1A. Consequently, DUS3L serves as a pivotal enzyme linking tRNA maturation pathways to the accuracy and efficiency of protein synthesis.
In the HGC-27 gastric cancer model, depletion of DUS3L is predicted to diminish dihydrouridine levels in tRNAs, leading to altered translation of proteins integral to cell proliferation, survival, and metastatic behavior. This disruption may uncover critical dependencies on translational fidelity for gastric tumor maintenance and progression. The polyclonal nature of the knockout product permits the study of varied editing efficiencies and phenotypic gradations, offering a physiologically relevant platform to dissect the contribution of tRNA modification defects to gastric adenocarcinoma pathophysiology.
This knockout product supports diverse research applications, including functional analysis of tRNA modifications in gastric cancer, investigation of translational control mechanisms in tumorigenesis, and preclinical evaluation of DUS3L as a therapeutic target. Recommended experimental approaches include tRNA modification profiling by liquid chromatography?Cmass spectrometry, ribosome profiling and RNA sequencing to assess translational efficiency, and standard cell-based assays for proliferation, colony formation, apoptosis, and migration/invasion. Expression analyses via Western blot and RT-qPCR can be used to confirm target disruption and explore downstream signaling consequences. For further details or to request a quote, please contact Ascent Research.
