A protein-independent fluorescent RNA aptamer reporter system for plant genetic engineering
Jiuyuan Bai, Yao Luo, Xin Wang, Li Shi, Mei Luo, Meng Yin, Yuanli Zuo, Guolin Li, Junyu Yao, Hua Yang, Mingdi Zhang, Wei Wei, Maolin Wang, Rui Wang, Chunhai Fan, Yun Zhao
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.