ArticleNature protocols2018
Single-mRNA detection in living S. cerevisiae using a re-engineered MS2 system.
Article in Nature protocols, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
16 citing papers in PubMed.
- Cyclin CLB2 mRNA localization and protein synthesis link cell cycle progression to bud growth.Nature communications · 2025Article
- Spatiotemporal regulation of endoplasmic reticulum stress visualized by live imaging of bipA mRNA in Aspergillus oryzae.Scientific reports · 2025Article
- Live cell imaging of β-tubulin mRNA reveals spatiotemporal expression dynamics in the filamentous fungus Aspergillus oryzae.Scientific reports · 2024Article
- A fast method to distinguish between fermentative and respiratory metabolisms in single yeast cells.iScience · 2024Article
- Intracellular RNA and DNA tracking by uridine-rich internal loop tagging with fluorogenic bPNA.Nature communications · 2023Article
- Avidity-based bright and photostable light-up aptamers for single-molecule mRNA imaging.Nature chemical biology · 2023Article
- Long-term imaging of individual mRNA molecules in living cells.Cell reports methods · 2022Article
- Intracellular mRNA transport and localized translation.Nature reviews. Molecular cell biology · 2021Review
- G3BPs in Plant Stress.Frontiers in plant science · 2021Review
- Live cell imaging and proteomic profiling of endogenous NEAT1 lncRNA by CRISPR/Cas9-mediated knock-in.Protein & cell · 2020Article
- Article
- Single molecule mRNA fluorescentData in brief · 2020Article
- New Generations of MS2 Variants and MCP Fusions to Detect Single mRNAs in Living Eukaryotic Cells.Methods in molecular biology (Clifton, N.J.) · 2020Article
- Imaging Single mRNA Molecules in Mammalian Cells Using an Optimized MS2-MCP System.Methods in molecular biology (Clifton, N.J.) · 2019Article
- Live-Cell Imaging of mRNP-NPC Interactions in Budding Yeast.Methods in molecular biology (Clifton, N.J.) · 2019Article
- Understanding spatiotemporal coupling of gene expression using single molecule RNA imaging technologies.TranscriptionArticle
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
3 authors.
Funding
Abstract
The MS2 system has been widely used, in organisms ranging from bacteria to higher eukaryotes, to image single mRNAs in intact cells with high precision. We have recently re-engineered the MS2 system for accurate detection of mRNAs in living Saccharomyces cerevisiae. Previous MS2 systems affected the degradation of the tagged mRNA, which led to accumulation of MS2 fragments and to erroneous conclusions about mRNA localization and expression. Here we describe a step-by-step protocol for the use of our latest MS2 system (MBSV6) for detecting endogenously tagged mRNAs using wide-field fluorescent microscopy in living yeast. The procedure is divided into three stages: tagging of endogenous gene with MBSV6 (~2 weeks), a two-color single-molecule RNA fluorescent in situ hybridization (smFISH) procedure to quantitatively assess whether mRNAs tagged with MS2 and MS2-coat protein (MCP) behave like untagged mRNAs (2 d, plus additional time for quantification), and a procedure to quantify single mRNAs by live imaging using wide-field microscopy (1 d, plus additional time for quantification). With this method it is now possible to interrogate all phases of mRNA expression, from transcription through decay. The described protocol is designed for S. cerevisiae; however, we think that our approach and the considerations discussed here can be extended to Escherichia coli, Drosophila, Caenorhabditis elegans, and mammalian cells.
Indexed as
Identifiers
30218101What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.