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

Jiuyuan BaiYao LuoXin WangShi LiMei LuoMeng YinYuanli ZuoGuolin LiJunyu YaoHua YangMingdi ZhangWei WeiMaolin WangRui WangChun-Hai FanYun Zhao
Published in: Nature communications (2020)
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.
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