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Trans-cis isomerization kinetics of cyanine dyes reports on the folding states of exogeneous RNA G-quadruplexes in live cells
| Title: | Trans-cis isomerization kinetics of cyanine dyes reports on the folding states of exogeneous RNA G-quadruplexes in live cells |
| Authors: | Kitamura, Akira Browse this author →KAKEN DB | | Tornmalm, Johan Browse this author | | Demirbay, Baris Browse this author | | Piguet, Joachim Browse this author | | Kinjo, Masataka Browse this author | | Widengren, Jerker Browse this author |
| Issue Date: | 21-Mar-2023 |
| Publisher: | Oxford University Press |
| Journal Title: | Nucleic acids research |
| Volume: | 51 |
| Issue: | 5 |
| Start Page: | e27 |
| End Page: | e27 |
| Publisher DOI: | 10.1093/nar/gkac1255 |
| Abstract: | Guanine (G)-rich nucleic acids are prone to assemble into four-stranded structures, so-called G-quadruplexes. Abnormal GGGGCC repeat elongations, and in particular their folding states, are associated with amyotrophic lateral sclerosis and frontotemporal dementia. Due to methodological constraints however, most studies of G quadruplex structures are restricted to in vitro conditions. Evidence of how GGGGCC repeats form into G-quadruplexes in vivo is sparse. We devised a readout strategy, exploiting the sensitivity of trans-cis isomerization of cyanine dyes to local viscosity and sterical constraints. Thereby, folding states of cyanine-labeled RNA, and in particular G-quadruplexes, can be identified in a sensitive manner. The isomerization kinetics, monitored via fluorescence blinking generated upon transitions between a fluorescent trans isomer and a non-fluorescent cis isomer, was first characterized for RNA with GGGGCC repeats in aqueous solution using fluorescence correlation spectroscopy and transient state (TRAST) monitoring. With TRAST, monitoring the isomerization kinetics from how the average fluorescence intensity varies with laser excitation modulation characteristics, we could then detect folding states of fluorescently tagged RNA introduced into live cells. |
| Type: | article |
| URI: | http://hdl.handle.net/2115/88907 |
| Appears in Collections: | 生命科学院・先端生命科学研究院 (Graduate School of Life Science / Faculty of Advanced Life Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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