In-Cell NMR Spectroscopy of Nucleic Acids in Human Cells

J 2019

In-Cell NMR Spectroscopy of Nucleic Acids in Human Cells

VÍŠKOVÁ, Pavlína, Daniel KRAFČÍK, Lukáš TRANTÍREK and Silvie TRANTÍRKOVÁ

Basic information

Original name

In-Cell NMR Spectroscopy of Nucleic Acids in Human Cells

Authors

VÍŠKOVÁ, Pavlína (203 Czech Republic, belonging to the institution), Daniel KRAFČÍK (703 Slovakia, belonging to the institution), Lukáš TRANTÍREK (203 Czech Republic, guarantor, belonging to the institution) and Silvie TRANTÍRKOVÁ (203 Czech Republic, belonging to the institution)

Edition

Current Protocols in Nucleic Acid Chemistry, United States, Wiley-Liss Inc, 2019, 1934-9270

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10609 Biochemical research methods

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

RIV identification code

RIV/00216224:14740/19:00108251

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1002/cpnc.71

Keywords in English

DNA; eukaryotic cellin-cell NMR spectroscopy; mammalian cell

Abstract

V originále

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.

Links

GA16-10504S, research and development project

Name: Charakterizace struktury nukleových kyselin v komplexním prostředí živých buněk pomocí vysoce rozlišené NMR spektroskopie

Investor: Czech Science Foundation

LQ1601, research and development project

Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

653706, interní kód MU

Name: iNEXT - Infrastructure for NMR, EM and X-ray crystallography for translational research (Acronym: iNEXT)

Investor: European Union, iNEXT - Infrastructure for NMR, EM and X-ray crystallography for translational research, RI Research Infrastructures (Excellent Science)

Citovat

VÍŠKOVÁ, Pavlína, Daniel KRAFČÍK, Lukáš TRANTÍREK and Silvie TRANTÍRKOVÁ. In-Cell NMR Spectroscopy of Nucleic Acids in Human Cells. Current Protocols in Nucleic Acid Chemistry. United States: Wiley-Liss Inc, 2019, vol. 76, No 1, p. 1-14. ISSN 1934-9270. Available from: https://dx.doi.org/10.1002/cpnc.71.

@article{1643298,
   author = {Víšková, Pavlína and Krafčík, Daniel and Trantírek, Lukáš and Trantírková, Silvie},
   article_location = {United States},
   article_number = {1},
   doi = {http://dx.doi.org/10.1002/cpnc.71},
   keywords = {DNA; eukaryotic cellin-cell NMR spectroscopy; mammalian cell},
   language = {eng},
   issn = {1934-9270},
   journal = {Current Protocols in Nucleic Acid Chemistry},
   title = {In-Cell NMR Spectroscopy of Nucleic Acids in Human Cells},
   url = {https://currentprotocols.onlinelibrary.wiley.com/doi/pdf/10.1002/cpnc.71},
   volume = {76},
   year = {2019}
}

TY  - JOUR
ID  - 1643298
AU  - Víšková, Pavlína - Krafčík, Daniel - Trantírek, Lukáš - Trantírková, Silvie
PY  - 2019
TI  - In-Cell NMR Spectroscopy of Nucleic Acids in Human Cells
JF  - Current Protocols in Nucleic Acid Chemistry
VL  - 76
IS  - 1
SP  - 1-14
EP  - 1-14
PB  - Wiley-Liss Inc
SN  - 19349270
KW  - DNA
KW  - eukaryotic cellin-cell NMR spectroscopy
KW  - mammalian cell
UR  - https://currentprotocols.onlinelibrary.wiley.com/doi/pdf/10.1002/cpnc.71
L2  - https://currentprotocols.onlinelibrary.wiley.com/doi/pdf/10.1002/cpnc.71
N2  - 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.
ER  -

VÍŠKOVÁ, Pavlína, Daniel KRAFČÍK, Lukáš TRANTÍREK and Silvie TRANTÍRKOVÁ. In-Cell NMR Spectroscopy of Nucleic Acids in Human Cells. \textit{Current Protocols in Nucleic Acid Chemistry}. United States: Wiley-Liss Inc, 2019, vol.~76, No~1, p.~1-14. ISSN~1934-9270. Available from: https://dx.doi.org/10.1002/cpnc.71.

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