In Cell NMR Spectroscopy: Investigation of G-Quadruplex Structures Inside Living Xenopus laevis Oocytes

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.