A fast and reliable method for monitoring genomic instability in the model organism Caenorhabditis elegans
| Autor: | Andrea Hartwig, Tanja Schwerdtle, Julia Bornhorst, Barbara Witt, Merle M. Nicolai |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Genomic instability
Life sciences biology Genome instability Health Toxicology and Mutagenesis ved/biology.organism_classification_rank.species Genotoxicity and Carcinogenicity Computational biology Toxicology Bleomycin chemistry.chemical_compound Alkaline unwinding tert-Butylhydroperoxide In vivo ddc:570 Animals Caenorhabditis elegans Model organism Organism Mutagenicity Tests ved/biology Reproducibility of Results General Medicine biology.organism_classification In vitro High-Throughput Screening Assays chemistry Cell culture DNA DNA Damage Mutagens |
| Zdroj: | Archives of Toxicology, 95, 3417–3424 Archives of Toxicology |
| ISSN: | 0340-5761 0003-9446 0370-8497 1432-0738 |
| DOI: | 10.5445/ir/1000137482 |
| Popis: | The identification of genotoxic agents and their potential for genotoxic alterations in an organism is crucial for risk assessment and approval procedures of the chemical and pharmaceutical industry. Classically, testing strategies for DNA or chromosomal damage focus on in vitro and in vivo (mainly rodent) investigations. In cell culture systems, the alkaline unwinding (AU) assay is one of the well-established methods for detecting the percentage of double-stranded DNA (dsDNA). By establishing a reliable lysis protocol, and further optimization of the AU assay for the model organism Caenorhabditis elegans (C. elegans), we provided a new tool for genotoxicity testing in the niche between in vitro and rodent experiments. The method is intended to complement existing testing strategies by a multicellular organism, which allows higher predictability of genotoxic potential compared to in vitro cell line or bacterial investigations, before utilizing in vivo (rodent) investigations. This also allows working within the 3R concept (reduction, refinement, and replacement of animal experiments), by reducing and possibly replacing animal testing. Validation with known genotoxic agents (bleomycin (BLM) and tert-butyl hydroperoxide (tBOOH)) proved the method to be meaningful, reproducible, and feasible for high-throughput genotoxicity testing, and especially preliminary screening. |
| Databáze: | OpenAIRE |
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