Citation:
He L, Binari R, Huang J, Falo-Sanjuan J, Perrimon N. In vivo study of gene expression with an enhanced dual-color fluorescent transcriptional timer. Elife. 2019;8.
2019_eLife_He.pdf | 4.29 MB | |
Supplement.zip | 58.47 MB |
Date Published:
2019 May 29Abstract:
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 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 . 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.See also: Research Article