Spectral recording of gene expression history by fluorescent timer protein

Monitoring spatio-temporal patterns of gene expression by fluorescent proteins requires longitudinal observation, which is often difficult to implement. Here, we fuse a fluorescent timer (FT) protein with an immediate early gene (IEG) promoter to track live gene expression in single cells. This resu...

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Veröffentlicht in:BioTechniques. - 1993. - 67(2019), 4 vom: 01. Okt., Seite 154-164
1. Verfasser: Tröscher, Anna R (VerfasserIn)
Weitere Verfasser: Werner, Barbara, Kaouane, Nadia, Haubensak, Wulf
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2019
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Journal Article Research Support, Non-U.S. Gov't flow cytometry fluorescent timer protein gene expression immediate early genes live cell imaging spatio-temporal resolution Luminescent Proteins Recombinant Fusion Proteins
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520 |a Monitoring spatio-temporal patterns of gene expression by fluorescent proteins requires longitudinal observation, which is often difficult to implement. Here, we fuse a fluorescent timer (FT) protein with an immediate early gene (IEG) promoter to track live gene expression in single cells. This results in a stimulus- and time-dependent spectral shift from blue to red for subsequent monitoring with fluorescence activated cell sorting (FACS) and live cell imaging. This spectral shift enables imputing the time point of activity post-hoc to dissociate early and late responders from a single snapshot in time. Thus, we provide a tool for tracking stimulus-driven IEG expression and demonstrate proof of concept exploiting promoter::FT fusions, adding new dimensions to experiments that require reconstructing spatio-temporal patterns of gene expression in cells, tissues or living organisms 
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650 4 |a Research Support, Non-U.S. Gov't 
650 4 |a flow cytometry 
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650 4 |a gene expression 
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650 4 |a live cell imaging 
650 4 |a spatio-temporal resolution 
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700 1 |a Kaouane, Nadia  |e verfasserin  |4 aut 
700 1 |a Haubensak, Wulf  |e verfasserin  |4 aut 
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