RRC ID 59155
Author He L, Binari R, Huang J, Falo-Sanjuan J, Perrimon N.
Title In vivo study of gene expression with an enhanced dual-color fluorescent transcriptional timer.
Journal Elife
Abstract 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.
Volume 8
Published 2019-5-29
DOI 10.7554/eLife.46181
PII 46181
PMID 31140975
PMC PMC6660218
MeSH Animals Drosophila melanogaster / embryology Drosophila melanogaster / genetics Embryo, Nonmammalian / cytology Embryo, Nonmammalian / metabolism Enhancer Elements, Genetic / genetics Fluorescence Gene Expression Regulation* Green Fluorescent Proteins / metabolism Intestines / cytology Protein Biosynthesis Receptors, Notch / metabolism STAT Transcription Factors / metabolism Stem Cells / cytology Transcription, Genetic*
IF 7.551
Times Cited 6
Resource
Drosophila DGRC#112553