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PMID: 23263691 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Single-molecule analysis of gene expression using two-color RNA labeling in live yeast.

Nature methods ·Vol. 10 ·No. 2 ·2013-02-00 ·Pages 119-21

Hocine S, Raymond P, Zenklusen D, Chao JA, Singer RH

Abstract

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.

MeSH Terms
ATPases Associated with Diverse Cellular Activities Adenosine Triphosphatases/genetics Gene Expression In Situ Hybridization, Fluorescence/methods Levivirus/genetics RNA Polymerase II/metabolism RNA, Fungal/metabolism RNA, Messenger/metabolism Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins/genetics
Chemicals
RNA, Fungal RNA, Messenger Saccharomyces cerevisiae Proteins RNA Polymerase II Adenosine Triphosphatases MDN1 protein, S cerevisiae ATPases Associated with Diverse Cellular Activities
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Hocine Sami
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York, USA.
Raymond Pascal
Zenklusen Daniel
Chao Jeffrey A
Singer Robert H
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Article Info
Journal
Nature methods
Abbr.
Nat Methods
ISSN
1548-7105
DOI
10.1038/nmeth.2305
Published
2013-02-00
Epub
2012-00-23
Pages
119-21
Language
English
Region
United States
NLM ID
101215604
PMCID
PMC3899799
Subset
IM
Grants
NIGMS NIH HHS · F32GM083430 · United States
NIGMS NIH HHS · R01 GM057071 · United States
NIGMS NIH HHS · GM57071 · United States
NIGMS NIH HHS · F32 GM083430 · United States
CIHR · 111099-1 · Canada
CIHR · MOP-BMB-232642 · Canada
External Links
PubMed source DOI: 10.1038/nmeth.2305 PMC Full Text View journal details
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