Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, 610065, China.
State Key Laboratory and Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, China.
Nat Commun. 2020 Jul 31;11(1):3847. doi: 10.1038/s41467-020-17497-7.
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.
报告基因系统在植物遗传工程和功能基因组学研究中被常规使用。大多数这样的植物报告基因系统会导致外源蛋白的积累。在这里，我们展示了一种基于荧光 RNA 适体的不依赖于蛋白质的报告基因系统，3WJ-4 × Bro。通过在大肠杆菌和本氏烟中进行瞬时表达试验，我们表明 3WJ-4 × Bro 适合于转基因鉴定和作为用于表达模式分析的 mRNA 报告基因。在拟南芥中稳定转化后，3WJ-4 × Bro 与靶转录物共分离和共表达，并通过多代稳定遗传。此外，3WJ-4 × Bro 可用于可视化植物中病毒介导的 RNA 传递。本研究展示了一种不依赖于蛋白质的报告基因系统，可用于转基因鉴定和 mRNA 的体内动态分析。
