In-Cell NMR Spectroscopy of Nucleic Acids in Human Cells

Curr Protoc Nucleic Acid Chem. 2019 Mar;76(1):e71. doi: 10.1002/cpnc.71. Epub 2018 Nov 29.

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.

Keywords: DNA, eukaryotic cell; in-cell NMR spectroscopy; mammalian cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Nucleus / metabolism
  • Cytosol / metabolism
  • Electroporation
  • Flow Cytometry
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Microscopy, Confocal
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Nucleic Acids / metabolism*
  • Transfection

Substances

  • Nucleic Acids