Real-Time Monitoring of G-Quadruplex Formation during Transcription

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• 作者：Tamaki Endoh ; Ambadas B. Rode ; Shuntaro Takahashi ; Yuka Kataoka ; Masayasu Kuwahara ; Naoki Sugimoto
• 刊名：Analytical Chemistry
• 出版年：2016
• 出版时间：February 16, 2016
• 年：2016
• 卷：88
• 期：4
• 页码：1984-1989
• 全文大小：460K
• ISSN：1520-6882

文摘

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.