In-Cell NMR Spectroscopy of Nucleic Acids in Human Cells

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Authors

VÍŠKOVÁ Pavlína KRAFČÍK Daniel TRANTÍREK Lukáš TRANTÍRKOVÁ Silvie

Year of publication 2019
Type Article in Periodical
Magazine / Source Current Protocols in Nucleic Acid Chemistry
MU Faculty or unit

Central European Institute of Technology

Citation
Web https://currentprotocols.onlinelibrary.wiley.com/doi/pdf/10.1002/cpnc.71
Doi http://dx.doi.org/10.1002/cpnc.71
Keywords DNA; eukaryotic cellin-cell NMR spectroscopy; mammalian cell
Description 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.
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