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

Nat Methods. 2013 Feb;10(2):119-21. doi: 10.1038/nmeth.2305. Epub 2012 Dec 23.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

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
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / genetics

Substances

  • RNA, Fungal
  • RNA, Messenger
  • Saccharomyces cerevisiae Proteins
  • RNA Polymerase II
  • Adenosine Triphosphatases
  • MDN1 protein, S cerevisiae
  • ATPases Associated with Diverse Cellular Activities