PacBio sequencing detects genome‐wide ultra‐low‐frequency substitution mutations resulting from exposure to chemical mutagens
Autor: | Vasily N. Dobrovolsky, Jaime A. Miranda, Javier R. Revollo |
---|---|
Rok vydání: | 2021 |
Předmět: |
Genetics
Alkylating Agents Mutation Mutagenicity Tests Epidemiology DNA damage Base pair Health Toxicology and Mutagenesis High-Throughput Nucleotide Sequencing Biology medicine.disease_cause Genome DNA sequencing chemistry.chemical_compound Phenotype chemistry Ethylnitrosourea Escherichia coli medicine Genetics (clinical) DNA Single molecule real time sequencing |
Zdroj: | Environmental and Molecular Mutagenesis. 62:438-445 |
ISSN: | 1098-2280 0893-6692 |
Popis: | Genetic toxicology uses several assays to identity mutagens and protects the public. Most of these assays, however, rely on reporter genes, can only measure mutation indirectly based on phenotype, and often require specific cell lines or animal models-features that impede their integration with existing and emerging toxicological models, such as organoids. In this study, we show that PacBio Single-Molecule, Real-Time (PB SMRT) sequencing identified substitution mutations caused by chemical mutagens in Escherichia coli by generating nearly error-free consensus reads after repeatedly inspecting both strands of circular DNA molecules. Using DNA from E. coli exposed to ethyl methanosulfonate (EMS) or N-ethyl-N-nitrosourea (ENU), PB SMRT sequencing detected mutation frequencies (MFs) and spectra comparable to those obtained by clone-sequencing from the same exposures. The optimized background MF of PB SMRT sequencing was ≤ 1 × 10-7 mutations per base pair (mut/bp). |
Databáze: | OpenAIRE |
Externí odkaz: |