In-Cell NMR Spectroscopy of Nucleic Acids in Human Cells

• Autoři: VÍŠKOVÁ Pavlína; KRAFČÍK Daniel; TRANTÍREK Lukáš; TRANTÍRKOVÁ Silvie
• Rok publikování: 2019
• Druh: Článek v odborném periodiku
• Časopis / Zdroj: Current Protocols in Nucleic Acid Chemistry
• Fakulta / Pracoviště MU: Středoevropský technologický institut
• www: https://currentprotocols.onlinelibrary.wiley.com/doi/pdf/10.1002/cpnc.71
• Doi: http://dx.doi.org/10.1002/cpnc.71
• Klíčová slova: DNA; eukaryotic cellin-cell NMR spectroscopy; mammalian cell
• Popis: 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.

Související projekty

• iNEXT - Infrastructure for NMR, EM and X-ray crystallography for translational research
• Charakterizace struktury nukleových kyselin v komplexním prostředí živých buněk pomocí vysoce rozlišené NMR spektroskopie
• CEITEC 2020