Combining Spinach-tagged RNA and gene localization to image gene expression in live yeast

• Published in: Nature communications 2015-11, Vol.6 (1), p.8882-8882, Article 8882
• Main Authors: Guet, David, Burns, Laura T., Maji, Suman, Boulanger, Jérôme, Hersen, Pascal, Wente, Susan R., Salamero, Jean, Dargemont, Catherine
• Format: Article
• Language: English
• Subjects: 14; 38; 631/1647/245; 631/208/199; 631/337/572; 631/80/389/2029; Aptamers, Nucleotide - chemistry; Aptamers, Nucleotide - genetics; Aptamers, Nucleotide - metabolism; Binding sites; Biochemistry, Molecular Biology; Biosynthesis; Cell Nucleus - genetics; Cell Nucleus - metabolism; Gene expression; Gene Expression Regulation, Fungal; Gene loci; Genomes; Genomics; Humanities and Social Sciences; Life Sciences; Localization; Microscopy; Molecular Imaging - methods; multidisciplinary; Proteins; RNA, Fungal - chemistry; RNA, Fungal - genetics; RNA, Fungal - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; Saccharomyces cerevisiae - chemistry; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Science; Science (multidisciplinary); Spinach; Transcription factors; Transcription, Genetic; Yeast
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms9882

Although many factors required for the formation of export-competent mRNPs have been described, an integrative view of the spatiotemporal coordinated cascade leading mRNPs from their site of transcription to their site of nuclear exit, at a single cell level, is still partially missing due to technological limitations. Here we report that the RNA Spinach aptamer is a powerful tool for mRNA imaging in live S. cerevisiae with high spatial-temporal resolution and no perturbation of the mRNA biogenesis properties. Dedicated image processing workflows are developed to allow detection of very low abundance of transcripts, accurate quantitative dynamic studies, as well as to provide a localization precision close to 100 nm at consistent time scales. Combining these approaches has provided a state-of-the-art analysis of the osmotic shock response in live yeast by localizing induced transcription factors, target gene loci and corresponding transcripts. Measuring single-cell mRNA dynamics is critical to understand gene expression. Here, using RNA Spinach technique to detect very low abundant mRNAs, Guet et al . report an analysis of the osmotic shock response in live yeast by localizing induced transcription factors, target gene loci and corresponding transcripts.