A protein-independent fluorescent RNA aptamer reporter system for plant genetic engineering

Jiuyuan Bai, Yao Luo, Xin Wang, Shi Li, Mei Luo, Meng Yin, Yuanli Zuo, Guolin Li, Junyu Yao, Hua Yang, Mingdi Zhang, Wei Wei, Maolin Wang, Rui Wang (), Chunhai Fan () and Yun Zhao ()
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Jiuyuan Bai: Sichuan University
Yao Luo: Sichuan University
Xin Wang: Sichuan University
Shi Li: Sichuan University
Mei Luo: Sichuan University
Meng Yin: Sichuan University
Yuanli Zuo: Sichuan University
Guolin Li: Sichuan University
Junyu Yao: Sichuan University
Hua Yang: Sichuan University
Mingdi Zhang: Sichuan University
Wei Wei: Sichuan University
Maolin Wang: Sichuan University
Rui Wang: Sichuan University
Chunhai Fan: Shanghai Jiao Tong University
Yun Zhao: Sichuan University

Nature Communications, 2020, vol. 11, issue 1, 1-14

Abstract: Abstract Reporter systems are routinely used in plant genetic engineering and functional genomics research. Most such plant reporter systems cause accumulation of foreign proteins. Here, we demonstrate a protein-independent reporter system, 3WJ-4 × Bro, based on a fluorescent RNA aptamer. Via transient expression assays in both Escherichia coli and Nicotiana benthamiana, we show that 3WJ-4 × Bro is suitable for transgene identification and as an mRNA reporter for expression pattern analysis. Following stable transformation in Arabidopsis thaliana, 3WJ-4 × Bro co-segregates and co-expresses with target transcripts and is stably inherited through multiple generations. Further, 3WJ-4 × Bro can be used to visualize virus-mediated RNA delivery in plants. This study demonstrates a protein-independent reporter system that can be used for transgene identification and in vivo dynamic analysis of mRNA.

Date: 2020
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DOI: 10.1038/s41467-020-17497-7

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