Whole-genome high-fidelity sequencing: A novel approach to detecting and characterization of mutagenicity in vivo

IF 2.3 4区 医学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Vasily N. Dobrovolsky , Tomonari Matsuda , Page McKinzie , Jaime Miranda , Javier R. Revollo
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引用次数: 0

Abstract

Direct DNA sequencing can be used for characterizing mutagenicity in simple and complex biological models. Recently we described a method of whole-genome sequencing for detecting mutations in simple models of cultured bacteria, mammalian cells, and nematode. In the current proof-of-concept study, we expand and improve our method for evaluating a more complex mammalian biological model in outbred mice. We detail the method by applying it to a small set of animals treated with a mutagen with known mutagenicity profiles, N-ethyl-N-nitrosourea (ENU), for consistency with the known data. Whole-genome high-fidelity sequencing (HiFi Sequencing) showed frequencies and spectra of background mutations in tissues of untreated mice that were consistent with normal ageing and characterized by spontaneous or enzymatic deamination of 5-methylcytosine. In mice treated with a single 40 mg/kg dose of ENU, the frequency of mutations in the genomic DNA of solid tissues increased up to 7-fold, with the greatest increase observed in the spleen and the smallest increase in the liver. The most common mutations detected in ENU-treated mice were T > A transitions and T > C transversions, consistent with the types of mutations caused by alkylating agents. The data suggest that HiFi Sequencing may be useful for characterizing mutagenicity of novel compounds in various biological models.

全基因组高保真测序:一种检测和表征体内致突变性的新方法。
直接DNA测序可用于表征简单和复杂生物模型中的致突变性。最近,我们描述了一种全基因组测序方法,用于检测培养细菌、哺乳动物细胞和线虫的简单模型中的突变。在目前的概念验证研究中,我们扩展和改进了我们的方法,以评估更复杂的哺乳动物生物学模型。为了与已知数据保持一致,我们将其应用于一小组用已知诱变特性的诱变剂N-乙基-N-亚硝基脲(ENU)处理的动物,详细介绍了该方法。全基因组高保真测序(HiFi测序)显示了未经治疗的小鼠组织中背景突变的频率和光谱,这些突变与正常衰老一致,并以5-甲基胞嘧啶的自发或酶促脱氨基为特征。在用单次40mg/kg剂量的ENU治疗的小鼠中,实体组织基因组DNA的突变频率增加了7倍,其中在脾脏中观察到最大的增加,在肝脏中观察到最小的增加。在ENU处理的小鼠中检测到的最常见的突变是T>A转换和T>C颠换,这与烷基化剂引起的突变类型一致。数据表明,HiFi测序可能有助于在各种生物模型中表征新化合物的致突变性。
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来源期刊
CiteScore
3.80
自引率
5.30%
发文量
84
审稿时长
105 days
期刊介绍: Mutation Research - Genetic Toxicology and Environmental Mutagenesis (MRGTEM) publishes papers advancing knowledge in the field of genetic toxicology. Papers are welcomed in the following areas: New developments in genotoxicity testing of chemical agents (e.g. improvements in methodology of assay systems and interpretation of results). Alternatives to and refinement of the use of animals in genotoxicity testing. Nano-genotoxicology, the study of genotoxicity hazards and risks related to novel man-made nanomaterials. Studies of epigenetic changes in relation to genotoxic effects. The use of structure-activity relationships in predicting genotoxic effects. The isolation and chemical characterization of novel environmental mutagens. The measurement of genotoxic effects in human populations, when accompanied by quantitative measurements of environmental or occupational exposures. The application of novel technologies for assessing the hazard and risks associated with genotoxic substances (e.g. OMICS or other high-throughput approaches to genotoxicity testing). MRGTEM is now accepting submissions for a new section of the journal: Current Topics in Genotoxicity Testing, that will be dedicated to the discussion of current issues relating to design, interpretation and strategic use of genotoxicity tests. This section is envisaged to include discussions relating to the development of new international testing guidelines, but also to wider topics in the field. The evaluation of contrasting or opposing viewpoints is welcomed as long as the presentation is in accordance with the journal''s aims, scope, and policies.
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