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{"title":"In-Cell NMR Spectroscopy of Nucleic Acids in Human Cells","authors":"Pavlina Viskova, Daniel Krafcik, Lukas Trantirek, Silvie Foldynova-Trantirkova","doi":"10.1002/cpnc.71","DOIUrl":null,"url":null,"abstract":"<p>In-cell NMR spectroscopy is a unique tool that enables the study of the structure and dynamics of biomolecules as well as their interactions in the complex environment of living cells at near-to-atomic resolution. In this article, detailed instructions are described for setting up an in-cell NMR experiment for monitoring structures of DNA oligonucleotides introduced into nuclei of living human cells via tailored electroporation. Detailed step-by-step protocols for both the preparation of an in-cell NMR sample as well as protocols for conducting essential control experiments including flow cytometry and confocal microscopy are described. The strengths and limitations of in-cell NMR experiments are discussed. © 2018 by John Wiley & Sons, Inc.</p>","PeriodicalId":10966,"journal":{"name":"Current Protocols in Nucleic Acid Chemistry","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2018-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpnc.71","citationCount":"7","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Nucleic Acid Chemistry","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpnc.71","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Chemistry","Score":null,"Total":0}
引用次数: 7
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Abstract
In-cell NMR spectroscopy is a unique tool that enables the study of the structure and dynamics of biomolecules as well as their interactions in the complex environment of living cells at near-to-atomic resolution. In this article, detailed instructions are described for setting up an in-cell NMR experiment for monitoring structures of DNA oligonucleotides introduced into nuclei of living human cells via tailored electroporation. Detailed step-by-step protocols for both the preparation of an in-cell NMR sample as well as protocols for conducting essential control experiments including flow cytometry and confocal microscopy are described. The strengths and limitations of in-cell NMR experiments are discussed. © 2018 by John Wiley & Sons, Inc.
人类细胞中核酸的细胞内核磁共振光谱
细胞内核磁共振波谱是一种独特的工具，可以研究生物分子的结构和动力学，以及它们在接近原子分辨率的活细胞复杂环境中的相互作用。在本文中，详细说明了建立一个细胞内核磁共振实验，以监测DNA寡核苷酸的结构，通过量身定制的电穿孔引入到活的人类细胞的细胞核。详细的一步一步的协议，无论是在细胞内核磁共振样品的制备，以及协议进行必要的控制实验，包括流式细胞术和共聚焦显微镜描述。讨论了细胞内核磁共振实验的优点和局限性。©2018 by John Wiley &儿子,Inc。
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