title: Single-molecule analysis of gene expression using two-color RNA labeling in live yeast.
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.
authors.affiliation: Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York, USA.
authors.forename: Sami
authors.initials: S
authors.lastname: Hocine
authors.forename: Pascal
authors.initials: P
authors.lastname: Raymond
authors.forename: Daniel
authors.initials: D
authors.lastname: Zenklusen
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Chao
authors.forename: Robert H
authors.initials: RH
authors.lastname: Singer
mesh_terms: D000074183:ATPases Associated with Diverse Cellular Activities; D000251:Adenosine Triphosphatases / Q000235:genetics; D015870:Gene Expression; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D017909:Levivirus / Q000235:genetics; D012319:RNA Polymerase II / Q000378:metabolism; D012331:RNA, Fungal / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
chemical_list: D012331:RNA, Fungal; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D012319:RNA Polymerase II; D000251:Adenosine Triphosphatases; C496538:MDN1 protein, S cerevisiae; D000074183:ATPases Associated with Diverse Cellular Activities