The DUS1L Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the human hepatocellular carcinoma Huh-7 cell line, engineered for targeted disruption of the DUS1L gene. This loss-of-function model enables investigation of tRNA dihydrouridine modification in liver cancer contexts without introducing defined clonal genetic alterations. The polyclonal format preserves cellular heterogeneity, offering a robust system for studying gene function in heterogeneous tumor cell populations.
The Huh-7 cell line was established from a hepatocellular carcinoma of a 57-year-old Japanese male and has become a cornerstone model in hepatocyte biology, hepatitis C virus (HCV) replication, and drug metabolism research. Huh-7 cells retain key hepatic features, including expression of liver-specific metabolic enzymes and susceptibility to viral infection, making them an ideal host for examining the interplay between tRNA modifications and hepatocarcinogenesis.
DUS1L encodes a tRNA dihydrouridine synthase that catalyzes the conversion of uridine to dihydrouridine in the D-loop of tRNA molecules, a modification critical for tRNA structural flexibility and translational fidelity. DUS1L functions downstream of mTORC1 and MYC signaling, integrating nutrient and stress cues to regulate protein synthesis. It interacts directly with tRNA substrates and ribosomes, and its activity modulates the translational elongation factors eEF1A and eEF2, thereby fine-tuning codon-specific translation efficiency and global protein output.
In Huh-7 cells, DUS1L knockout is expected to perturb the dihydrouridine landscape of tRNAs, leading to altered translational elongation dynamics and potential shifts in the proteome under both basal and stress conditions. This disruption may impact cellular stress responses and oncogenic phenotypes such as proliferation, migration, and drug sensitivity, providing a platform to dissect how translational dysregulation contributes to hepatocellular carcinoma progression.
This knockout product is suited for a range of experimental applications, including tRNA sequencing to map dihydrouridine modifications, polysome profiling and puromycin incorporation assays to measure translation elongation rates, and cell viability or colony formation assays to evaluate growth phenotypes. The model can also be used in drug sensitivity testing and synthetic lethality screens to identify vulnerabilities in DUS1L-deficient liver cancer cells. For additional technical information, please contact Ascent Research.
