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

eLife

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[1] Department of Genetics, Harvard Medical School, Boston, United States;Department of Genetics, Harvard Medical School, Boston, United States;Howard Hughes Medical Institute, Boston, United States;International Academy of Targeted Therapeutics and Innovation, Chongqing University of Arts and Sciences, Chongqing, China;Tufts University, Medford, United States;

关键词: transcriptional dynamics;    fluorescent reporter;    transcriptional timer;    D. melanogaster;   

DOI  :  10.7554/eLife.46181

来源: publisher

【 摘 要 】

10.7554/eLife.46181.001Fluorescent 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.

【 授权许可】

CC BY