Protocol

Imaging Gene Expression

Volume 1042 of the series Methods in Molecular Biology pp 101-113

Date:

Imaging Nascent RNA Dynamics in Dictyostelium

  • Jonathan R. ChubbAffiliated withMRC Laboratory for Molecular Cell Biology, Department of Cell and Developmental Biology, University College London
  • , Michelle StevenseAffiliated withInstitut für Physiologische Chemie, Medizinische Fakultät, TU Dresden
  • , Danielle CannonAffiliated withDepartment of Cell and Developmental Biology and MRC Laboratory for Molecular Cell Biology, University College London
  • , Tetsuya MuramotoAffiliated withLaboratory for Cell Signaling Dynamics, Quantitative Biology Center, RIKEN
  • , Adam M. CorriganAffiliated withMRC Laboratory for Molecular Cell Biology, Department of Cell and Developmental Biology, University College London

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Abstract

Dictyostelium cells have great utility for live imaging of single gene transcriptional dynamics. The cells allow efficient molecular genetics, for targeting of RNA reporters and fluorescent proteins to individual, defined loci. Dictyostelium cells share many signalling, chromatin and nuclear characteristics of larger eukaryotes, yet the cells have a relatively simple scattered differentiation programme, allowing imaging of transcriptional events in the context of stochastic developmental choices. This review will detail the methods and considerations for imaging nascent RNA dynamics at single genes in living Dictyostelium cells.

Key words

Chromatin Dictyostelium Morphogen Live-cell imaging Transcriptional bursting Pulsing Stochastic gene expression Transcriptional noise Transcriptional pausing