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

Nat Commun. 2020 Jul 31;11(1):3847. doi: 10.1038/s41467-020-17497-7.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aptamers, Nucleotide / chemistry
  • Aptamers, Nucleotide / genetics*
  • Aptamers, Nucleotide / metabolism
  • Arabidopsis / genetics*
  • Arabidopsis / metabolism
  • Benzyl Compounds / chemistry
  • Brassica / genetics*
  • Brassica / metabolism
  • Fluorescence
  • Fluorescent Dyes / chemistry
  • Gene Expression Regulation
  • Genes, Reporter
  • Genetic Engineering / methods*
  • Imidazolines / chemistry
  • Nicotiana / genetics*
  • Nicotiana / metabolism
  • Plant Leaves / genetics
  • Plant Leaves / metabolism
  • Plants, Genetically Modified
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • Transformation, Genetic

Substances

  • 3,5-difluoro-4-hydroxybenzylidene imidazolinone
  • Aptamers, Nucleotide
  • Benzyl Compounds
  • Fluorescent Dyes
  • Imidazolines
  • RNA, Messenger