The DTD1 Knockout A2780 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A2780 human ovarian carcinoma cell line, carrying a targeted disruption of the DTD1 gene. This loss-of-function model is generated using CRISPR/Cas9-mediated gene disruption to ablate DTD1 expression without introducing a defined clonal selection step, resulting in a heterogeneous pool of edited cells suitable for diverse functional assays.
The parental A2780 cell line is a widely used epithelial model of human ovarian carcinoma, originally established from an untreated patient. These adherent cells retain key molecular features of ovarian cancer, including relevant oncogenic signaling pathways and drug response profiles, making them an invaluable tool for cancer biology research, particularly in studies of chemoresistance, tumor progression, and the identification of novel therapeutic vulnerabilities.
DTD1 encodes D-aminoacyl-tRNA deacylase, an enzyme that hydrolyzes D-aminoacyl-tRNAs generated by mischarging by aminoacyl-tRNA synthetases. This activity prevents D-amino acid incorporation into proteins, maintaining translational fidelity at the ribosome. DTD1 physically interacts with tRNA and acts downstream of aminoacyl-tRNA synthetase function to ensure proteome integrity. Its role in D-amino acid metabolism is critical for cellular resistance to D-amino acid toxicity.
In the A2780 ovarian cancer context, loss of DTD1 function disrupts this protective deacylation activity, leading to the accumulation of D-aminoacyl-tRNAs and subsequent misincorporation of D-amino acids into proteins. This renders the cells exquisitely sensitive to D-amino acid-induced stress, providing a powerful platform to investigate the consequences of translational infidelity in a cancer model. Given the altered metabolic state of cancer cells, this knockout model may reveal unique vulnerabilities related to proteostasis and amino acid metabolism that are not apparent in non-transformed cells.
Researchers can use these polyclonal knockout cells to study translational quality control and D-amino acid toxicity. Viability assays with D-amino acids assess cytotoxicity, while mass spectrometry detects D-amino acid incorporation. Western blotting confirms DTD1 disruption. Protein synthesis and colony formation assays evaluate translational fidelity and long-term survival. This model enables screening of compounds targeting D-amino acid metabolism or translational machinery in ovarian cancer. For further information, contact Ascent Research.
