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Spinach-based RNA mimicking GFP in plant cells

Yu, Z., Wang, Y., Mei, F., Yan, H., Jin, Zhenhui, Zhang, Pengcheng, Zhang, X., Tor, Mahmut ORCID: https://orcid.org/0000-0002-4416-5048, Jackson, S., Shi, N. and Hong, Yiguo (2022) Spinach-based RNA mimicking GFP in plant cells. Functional & Integrative Genomics. ISSN Online: 1438-7948

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Abstract

Spinach RNA-mimicking GFP (S-RMG) has been successfully used to monitor cellular RNAs including microRNAs in bacterium, yeast, and human cells. However, S-RMG has not been established in plants. In this study, we found that like bacterial, yeast, and human cellular tRNAs, plant tRNAs such as tRNALys can protect and/or stabilize the Spinach RNA aptamer interaction with the fluorophore DFHBI enabling detectable levels of green fluorescence to be emitted. The tRNALys-Spinach-tRNALys, once delivered into “chloroplast-free” onion epidermal cells can emit strong green fluorescence in the presence of DFHBI. Our results demonstrate for the first time that Spinach-based RNA visualization has the potential for in vivo monitoring of RNAs in plant cells.

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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/.

This work was in part supported by grants from the Ministry of Science & Technology of China (National Key R&D Program 2017YFE0110900), Ministry of Agriculture of the People’s Republic of China (National Transgenic Program of China 2016ZX08009001-004), National Natural Science Foundation of China (31200913, 31872636), Zhejiang Provincial Natural Science Foundation (LY19C020002), China Scholarship Council (201709645003), the Entrepreneurship and Innovation Project for the Overseas Returnees (or Teams) in Hangzhou (4105C5062000611), Hangzhou Normal University (Sino-EU Plant RNA Signaling S&T Platform Initiative 9995C5021841101), and the UK BBSRC China Partnering Awards (BB/T018295/1).

Uncontrolled Discrete Keywords: Onion epidermal cell, RNA aptamer, RNA fluorescence, Spinach-RMG
Divisions: College of Health, Life and Environmental Sciences > School of Science and the Environment
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Copyright Info: © The Author(s) 2022
Depositing User: Mahmut Tor
Date Deposited: 11 Mar 2022 15:55
Last Modified: 11 Mar 2022 16:04
URI: https://eprints.worc.ac.uk/id/eprint/11757

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