| Hyperactive DNA Cutting for Unbiased UHPLC-MS/MS Quantification of Epigenetic DNA Marks by Engineering DNase I Mutants | |
Song, Xingrui; Song, Xinyue; Lai, Weiyi ; Wang, Hailin
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| 2023 | |
| Source Publication | ANALYTICAL CHEMISTRY
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| ISSN | 0003-2700 |
| Pages | 1-7 |
| Abstract | Epigenetic DNA modifications, such as 5-methylcytosine, 5-hydroxymethylcytosine, and 5-formylcytosine, are associated with a variety of diseases and potential biomarkers for cancer diagnosis and therapy. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) assays are considered to be the gold standard for qualitative and quantitative detection of DNA modifications. DNA digestion for converting long DNA polymer into 2 '-deoxynucleosides is an important preprocessing step to achieve sensitive and accurate LC-MS/MS quantification. Here, we showed that, as stimulated by divalent metal ions, Mg2+ and Mn2+, the engineered human DNase I Q9R:E13R:N74K mutant can efficiently digest DNA in the presence of monovalent metal ions at a high concentration (e.g., 1 M NaCl), showing hyperactivity on DNA cutting. We also found that the engineered DNase I mutants display exceptional DNA-cutting activity over a wider pH range (5.5-9.5). Due to their hyperactivity and high salt tolerance, the engineered DNase I mutants cut DNA 5mC and dC efficiently. Benefitting from this DNA-cutting hyperactivity, we demonstrated an LC-MS/MS assay for unbiased and accurate quantification of DNA 5mC. |
| Department | 环境化学与生态毒理学国家重点实验室 |
| Document Type | 期刊论文 |
| Identifier | https://ir.rcees.ac.cn/handle/311016/48493 |
| Collection | 环境化学与生态毒理学国家重点实验室 |
| Affiliation | 1.Chinese Academy of Sciences 2.University of Chinese Academy of Sciences, CAS 3.Research Center for Eco-Environmental Sciences (RCEES) |
| Recommended Citation GB/T 7714 | Song, Xingrui,Song, Xinyue,Lai, Weiyi,et al. Hyperactive DNA Cutting for Unbiased UHPLC-MS/MS Quantification of Epigenetic DNA Marks by Engineering DNase I Mutants[J]. ANALYTICAL CHEMISTRY,2023:1-7. |
| APA | Song, Xingrui,Song, Xinyue,Lai, Weiyi,&Wang, Hailin.(2023).Hyperactive DNA Cutting for Unbiased UHPLC-MS/MS Quantification of Epigenetic DNA Marks by Engineering DNase I Mutants.ANALYTICAL CHEMISTRY,1-7. |
| MLA | Song, Xingrui,et al."Hyperactive DNA Cutting for Unbiased UHPLC-MS/MS Quantification of Epigenetic DNA Marks by Engineering DNase I Mutants".ANALYTICAL CHEMISTRY (2023):1-7. |
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| Hyperactive DNA Cutt（3972KB） | 期刊论文 | 出版稿 | 开放获取 | CC BY-NC-SA | View Download | |
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