In vivo study of gene expression with an enhanced dual-color fluorescent transcriptional timer.

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Binari, Richard 
Huang, Jiuhong 
Falo-Sanjuan, Julia 

Fluorescent transcriptional reporters are widely used as signaling reporters and biomarkers to monitor pathway activities and determine cell type identities. However, a large amount of dynamic information is lost due to the long half-life of the fluorescent proteins. To better detect dynamics, fluorescent transcriptional reporters can be destabilized to shorten their half-lives. However, applications of this approach in vivo are limited due to significant reduction of signal intensities. To overcome this limitation, we enhanced translation of a destabilized fluorescent protein and demonstrate the advantages of this approach by characterizing spatio-temporal changes of transcriptional activities in Drosophila. In addition, by combining a fast-folding destabilized fluorescent protein and a slow-folding long-lived fluorescent protein, we generated a dual-color transcriptional timer that provides spatio-temporal information about signaling pathway activities. Finally, we demonstrate the use of this transcriptional timer to identify new genes with dynamic expression patterns.

D. melanogaster, developmental biology, fluorescent reporter, transcriptional dynamics, transcriptional timer, Animals, Drosophila melanogaster, Embryo, Nonmammalian, Enhancer Elements, Genetic, Fluorescence, Gene Expression Regulation, Green Fluorescent Proteins, Intestines, Protein Biosynthesis, Receptors, Notch, STAT Transcription Factors, Stem Cells, Transcription, Genetic
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eLife Sciences Publications, Ltd