The DUS1L Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the DUS1L gene in the human 786-O renal carcinoma epithelial cell line. This product consists of a diverse pool of cells carrying various DUS1L alleles generated by CRISPR/Cas9-mediated gene targeting, offering a population-level loss-of-function model that avoids the phenotypic artifacts associated with single-cell clones. The polyclonal format is particularly suited for high-throughput screening, pooled functional genomics, and assays requiring robust statistical averaging across genetic variants.
The parental 786-O cell line is derived from a primary clear cell renal cell carcinoma (ccRCC) and harbors a biallelic mutation in the von Hippel-Lindau (VHL) tumor suppressor gene. This defect leads to constitutive stabilization of hypoxia-inducible factors (HIF1??/2??), driving aberrant transcription of genes involved in angiogenesis, glycolysis, and cell growth. Additionally, VHL loss potentiates mTORC1 signaling, a master regulator of protein synthesis. These molecular features establish 786-O as a highly relevant model for investigating translational control mechanisms in ccRCC.
DUS1L encodes a tRNA dihydrouridine synthase that catalyzes the conversion of uridine to dihydrouridine in tRNAs, promoting translation fidelity and efficiency. The enzyme is regulated by MYC and mTORC1 signaling, linking its function to cellular growth pathways. DUS1L acts on tRNA substrates, interacting with translation elongation factors and ribosomes to modulate codon-dependent protein synthesis. Knockout of DUS1L impairs dihydrouridine modification, potentially reducing the translation of mRNAs with specific codon usage, thereby affecting the proteome.
In the 786-O ccRCC background, DUS1L knockout provides a precise tool to explore how tRNA modifications contribute to the dysregulated translation programs driven by VHL loss and consequent MYC/mTORC1 hyperactivation. Since tumor cells often rely on elevated translational capacity to sustain rapid growth, disruption of DUS1L may render ccRCC cells susceptible to inhibitors of the translational apparatus. This model thus enables the investigation of synthetic lethal relationships and the identification of therapeutic targets within the tRNA modification axis of renal carcinoma.
Researchers can employ this knockout model for detailed functional studies, including western blotting to confirm DUS1L ablation, puromycin incorporation assays to measure global protein synthesis, and ribosome profiling to assess codon-specific translational changes. Complementary assays such as mass spectrometry-based dihydrouridine quantification, cell proliferation (MTT/BrdU), cell cycle analysis, and colony formation allow comprehensive phenotypic characterization. These approaches facilitate research into renal cancer biology, tRNA epitranscriptomics, and translational regulation. For further technical information, please contact Ascent Research.
