Journal
ANALYTICAL CHEMISTRY
Volume 88, Issue 4, Pages 1984-1989Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.5b04396
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Funding
- MEXT (Japan)
- Hirao Taro Foundation of KONAN GAKUEN for Academic Research
- Grants-in-Aid for Scientific Research [26350964, 26620138] Funding Source: KAKEN
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Cotranscriptional folding of an RNA transcript enables formation of metastable RNA structures. Thermodynamic and kinetic properties of RNA G-quadruplex formation have previously been investigated using purified guanine-rich oligonucleotides. Here, we describe a method for analysis of cotranscriptional dynamics of the G-quadruplex formation based on real-time monitoring of the fluorescence of G-quadruplex ligands. For RNA sequences with the potential to form mutually exclusive hairpin or G-quadruplex structures, the efficiency of G-quadruplex formation during transcription depended on position of the hairpin forming sequence. The real-time monitoring enabled evaluation of environmental effects on RNA dynamics, as we demonstrated facilitation of post-transcriptional G-quadruplex formation under molecular crowding conditions. The strategy demonstrated here provides folding insights into the G-quadruplex during transcription that should be involved in gene regulation.
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