Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/151981
Title: Photophysics of DFHBI bound to RNA aptamer baby spinach
Authors: Dao, Nguyen Thuan
Haselsberger, Reinhard
Khuc, Mai-Thu
Phan, Anh Tuân
Voityuk, Alexander A.
Michel-Beyerle, Maria-Elisabeth
Keywords: Science::Biological sciences
Issue Date: 2021
Source: Dao, N. T., Haselsberger, R., Khuc, M., Phan, A. T., Voityuk, A. A. & Michel-Beyerle, M. (2021). Photophysics of DFHBI bound to RNA aptamer baby spinach. Scientific Reports, 11(1), 7356-. https://dx.doi.org/10.1038/s41598-021-85091-y
Journal: Scientific Reports
Abstract: The discovery of the GFP-type dye DFHBI that becomes fluorescent upon binding to an RNA aptamer, termed Spinach, led to the development of a variety of fluorogenic RNA systems that enable genetic encoding of living cells. In view of increasing interest in small RNA aptamers and the scarcity of their photophysical characterisation, this paper is a model study on Baby Spinach, a truncated Spinach aptamer with half its sequence. Fluorescence and fluorescence excitation spectra of DFHBI complexes of Spinach and Baby Spinach are known to be similar. Surprisingly, a significant divergence between absorption and fluorescence excitation spectra of the DFHBI/RNA complex was observed on conditions of saturation at large excess of RNA over DFHBI. Since absorption spectra were not reported for any Spinach-type aptamer, this effect is new. Quantitative modelling of the absorption spectrum based on competing dark and fluorescent binding sites could explain it. However, following reasoning of fluorescence lifetimes of bound DFHBI, femtosecond-fluorescence lifetime profiles would be more supportive of the notion that the abnormal absorption spectrum is largely caused by trans-isomers formed within the cis-bound DFHBI/RNA complex. Independent of the origin, the unexpected discrepancy between absorption and fluorescence excitation spectra allows for easily accessed screening and insight into the efficiency of a fluorogenic dye/RNA system.
URI: https://hdl.handle.net/10356/151981
ISSN: 2045-2322
DOI: 10.1038/s41598-021-85091-y
Rights: © 2021 The Author(s). Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SPMS Journal Articles

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