Single-molecule analysis of gene expression using two-color RNA labeling in live yeast
Open accessAbstract
Live-cell imaging of mRNA yields important insights into gene expression, but it has generally been limited to the labeling of one RNA species and has never been used to count single mRNAs over time in yeast. We demonstrate a two-color imaging system with single-molecule resolution using MS2 and PP7 RNA labeling. We use this methodology to measure intrinsic noise in mRNA levels and RNA polymerase II kinetics at a single gene.
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Using constructs carrying arrays of MS2 and PP7 repeats in the 5′ and 3′ regions of the same gene, RNA Pol II speed was estimated to range from 0.84 kb/min to 3.66 kb/min.
“By using constructs carrying arrays of MS2 and PP7 repeats in the 5′ and 3′ regions of the same gene, RNA Pol II speed was estimated to range from 0.84 kb/min to 3.66 kb/min (Hocine et al, 2013; Fukaya et al, 2017).”
From RNA polymerase II speed: a key player in controlling and adapting transcriptome composition (2021) · cites this paper as Hocine et al, 2013
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- Authors
- Hocine, S., Raymond, P., Zenklusen, D., Chao, J. A., & Singer, R. H.
- Affiliations
- Albert Einstein College of Medicine (US)
- Université de Montréal (CA)
- Corresponding author
- Singer, Robert H
- Venue
- Nature Methods
- Published
- 2012-12-23
- Volume
- 10
- Issue
- 2
- Pages
- 119-121
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- PMC
- PMC3899799
- OpenAlex
- W2009743304
- Cited by
- 297
- References
- 22
- Citations by year
- 2026: 8 · 2025: 10 · 2024: 20 · 2023: 16 · 2022: 17 · 2021: 27 · 2020: 24 · 2019: 22 · 2018: 31 · 2017: 26 · 2016: 29 · 2015: 36 · 2014: 22 · 2013: 9
- FWCI
- 9.79
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Methods (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Canadian Institutes of Health Research (MOP-BMB-232642, 111099_1)
- National Institute of General Medical Sciences (F32GM083430, R01GM057071, GM57071)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene Regulatory Network Analysis
- Keywords
- single-molecule RNA imaging, MS2, live-cell imaging, yeast
- MeSH terms
- ATPases Associated with Diverse Cellular Activities, Adenosine Triphosphatases, RNA Polymerase II, RNA, Fungal, RNA, Messenger, Saccharomyces cerevisiae, Gene Expression, In Situ Hybridization, Fluorescence, Levivirus, Saccharomyces cerevisiae Proteins
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