In collaboration with the research team of Professor Chunhai Fan of Shanghai Jiaotong University, Professor Yun Zhao’s team from the College of Life Sciences, Sichuan University has recently published a paper entitled “A Protein-independent Fluorescent RNA Aptamer Reporter System for Plant Genetic Engineering” in Nature Communications online. In this research, the researchers combined nanotechnology with biotechnology, and constructed a reporter system of plant genetic transformation at RNA level for the first time, which was verified in Arabidopsisthaliana and tobacco genetic transformation . The first authors are doctoral students Jiuyuan Bai, Yao Luo and Xin Wang of the College of Life Sciences. Prof. Yun Zhao is the corresponding author and Sichuan University is the first work unit of the paper.

“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.” (Abstract)

Fig. One: Comparison of 3WJ-nBro series for imaging mRNA in vitro and in vivo

Fig. 2:Performance of 3WJ-4 × Bro in E. coli cells for reporting the expression of various genes

Fig. 3: Identification of transgenic A. thaliana and expression analysis of target genes using the 3WJ-4 × Bro/DFBH-1T reporter system

This project is the first of its kind to construct a plant genetic engineering reporter system at RNA level through nucleic acid nanotechnology, which is an important achievement by virtue of the intersection of nanoscience and life science, and breaks through the application limitation of protein reporter system. At the same time, it also provides a new visual tool for RNA localization, behavior and function research, which is of great significance for the development and application of plant genetic engineering

Article link: https://www.nature.com/articles/s41467-020-17497-7