Enzymatic incorporation of an isotope-labeled adenine into RNA for the study of conformational dynamics by NMR.

ABSTRACT: Solution NMR spectroscopy is a well-established tool with unique advantages for structural studies of RNA molecules. However, for large RNA sequences, the NMR resonances often overlap severely. A reliable way to perform resonance assignment and allow further analysis despite spectral crowding is the use of site-specific isotope labeling in sample preparation. While solid-phase oligonucleotide synthesis has several advantages, RNA length and availability of isotope-labeled building blocks are persistent issues. Purely enzymatic methods represent an alternative and have been presented in the literature. In this study, we report on a method in which we exploit the preference of T7 RNA polymerase for nucleotide monophosphates over triphosphates for the 5' position, which allows 5'-labeling of

SUBMITTER: Feyrer H 

PROVIDER: S-EPMC9269771 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature