Abstract
G-quadruplexes are inherently polymorphic nucleic acid structures. Their folding topology depends on the nucleic acid primary sequence and on physical–chemical environmental factors. Hence, it remains unclear if a G-quadruplex topology determined in the test tube (in vitro) will also form in vivo. Characterization of G-quadruplexes in their native environment has been proposed as an efficient strategy to tackle this issue. So far, characterization of G-quadruplex structures in living cells has relied exclusively on the use of Xenopus laevis oocytes as a eukaryotic cell model system. Here, we describe the protocol for the preparation of X. laevis oocytes for studies of G-quadruplexes as well as other nucleic acids motifs under native conditions using in-cell NMR spectroscopy.
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Acknowledgments
This work was supported by grants from the Czech Science Foundation (17-12075S), European Regional Development Fund (SYMBIT:Â CZ.02.1.01/0.0/0.0/15_003/0000477), Horizon 2020 Program of the EU (iNEXT: grant agreement 653706), and from the MEYS CR (CEITEC 2020 LQ1601).
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Krafcikova, M., Hänsel-Hertsch, R., Trantirek, L., Foldynova-Trantirkova, S. (2019). In Cell NMR Spectroscopy: Investigation of G-Quadruplex Structures Inside Living Xenopus laevis Oocytes. In: Yang, D., Lin, C. (eds) G-Quadruplex Nucleic Acids. Methods in Molecular Biology, vol 2035. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9666-7_25
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DOI: https://doi.org/10.1007/978-1-4939-9666-7_25
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