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Methods Mol Biol. 2018;1649:349-357. doi: 10.1007/978-1-4939-7213-5_23.

Live Imaging of mRNA Synthesis in Drosophila.

Author information

1
Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.
2
Department of Physics, and Biophysics Graduate Group, University of California, Berkeley, CA, USA.
3
Department of Physics, Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA. tg2@princeton.edu.

Abstract

mRNA synthesis is one of the earliest readouts of the activity of a transcribed gene, which is of particular interest in the context of metazoan cell fate specification. These processes are intrinsically dynamic and stochastic, which makes in vivo single-cell measurements inevitable. Here, we present the application of a technology that has been widely used in single celled organisms to measure transcriptional activity in developing embryos of the fruit fly Drosophila melanogaster. The method allows for quantification of instantaneous polymerase occupancy of active gene loci and thereby enables the development and testing of models of gene regulation in development.

KEYWORDS:

Drosophila; Fluorescence; Nascent mRNA; Quantitative live imaging; Transcription activity

PMID:
29130209
PMCID:
PMC5909709
DOI:
10.1007/978-1-4939-7213-5_23
[Indexed for MEDLINE]
Free PMC Article

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