Monitoring dynamics of single-cell gene expression over multiple cell cycles

Mol Syst Biol. 2005:1:2005.0024. doi: 10.1038/msb4100032. Epub 2005 Nov 22.

Abstract

Recent progress in reconstructing gene regulatory networks has established a framework for a quantitative description of the dynamics of many important cellular processes. Such a description will require novel experimental techniques that enable the generation of time-series data for the governing regulatory proteins in a large number of individual living cells. Here, we utilize microfabrication to construct a Tesla microchemostat that permits single-cell fluorescence imaging of gene expression over many cellular generations. The device is used to capture and constrain asymmetrically dividing or motile cells within a trapping region and to deliver nutrients and regulate the cellular population within this region. We illustrate the operation of the microchemostat with Saccharomyces cerevisiae and explore the evolution of single-cell gene expression and cycle time as a function of generation. Our findings highlight the importance of novel assays for quantifying the dynamics of gene expression and cellular growth, and establish a methodology for exploring the effects of gene expression on long-term processes such as cellular aging.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacterial Proteins / analysis
  • Cell Cycle
  • Cell Movement
  • Equipment Design
  • Gene Expression Profiling / instrumentation
  • Gene Expression Profiling / methods*
  • Gene Expression*
  • Luminescent Proteins / analysis
  • Microchemistry
  • Microscopy, Fluorescence / instrumentation*
  • Microscopy, Fluorescence / methods
  • Microspheres
  • Mycology / instrumentation
  • Mycology / methods
  • Recombinant Fusion Proteins / biosynthesis
  • Red Fluorescent Protein
  • Rheology
  • Saccharomyces cerevisiae / cytology
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / biosynthesis*
  • Saccharomyces cerevisiae Proteins / genetics
  • Software

Substances

  • Bacterial Proteins
  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • yellow fluorescent protein, Bacteria