The DUS3L Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population for functional investigation of DUS3L. This product is generated through CRISPR/Cas9-mediated gene disruption of the DUS3L locus in K-562 cells, resulting in a mixed population of cells carrying loss-of-function mutations. The polyclonal format preserves genetic diversity, allowing study of collective DUS3L knockout effects without single-cell cloning biases. This model is tailored for exploring DUS3L’s role in tRNA modification, translation regulation, and leukemia cell biology.
The host cell line, K-562, is a human chronic myeloid leukemia (CML) cell line derived from the pleural effusion of a 53-year-old female patient in blast crisis. K-562 cells are Philadelphia chromosome-positive myeloid progenitors, expressing BCR-ABL and serving as a leukemia model. The leukemic background provides a pathologically relevant context for examining how DUS3L-dependent tRNA modifications influence cancer cell behavior, including growth, apoptosis, and stress responses.
DUS3L encodes a dihydrouridine synthase that catalyzes the reduction of uridine to dihydrouridine in tRNA molecules, a modification critical for tRNA stability and translational fidelity. DUS3L functions alongside its paralog DUS3 and interacts directly with tRNA substrates and the elongator complex. The enzymatic activity modulates the dihydrouridine landscape of the tRNA pool, influencing protein synthesis efficiency and accuracy. In this model, disrupted DUS3L activity may lead to altered translation of specific mRNA transcripts, particularly those with codon usage dependent on modified tRNAs. While upstream regulators remain uncharacterized, downstream effects converge on the translation machinery, impacting fundamental cellular processes.
Knocking out DUS3L in K-562 leukemia cells allows direct interrogation of how tRNA modification pathways contribute to malignancy. Changes in dihydrouridine levels can rewire the proteome by selectively affecting translation of proteins involved in cell cycle regulation, apoptosis, and metabolic adaptation. The polyclonal knockout population is ideal for pooled phenotypic screens, assessing global changes in proliferation, drug sensitivity, and stress resilience. It also enables synthetic lethal interaction studies with other tRNA modification factors, potentially revealing new targets in leukemia. The leukemic background ensures translational relevance.
Typical applications include tRNA sequencing for modification profiling, polysome profiling and ribosome footprinting for translation analysis, and proliferation/apoptosis assays. Western blotting and RT-qPCR can quantify changes in translation-related proteins and tRNA levels. These cells are suitable for high-throughput screens to identify DUS3L pathway modulators. For further information, contact Ascent Research.
