Spectral recording of gene expression history by fluorescent timer protein

BioTechniques

Nadia Kaouane¹  Anna R Tröscher¹  Barbara Werner¹  Wulf Haubensak¹ 

[1] 1Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC), Campus Vienna Biocenter 1, 1030 Vienna, Austria;

关键词: flow cytometry;    fluorescent timer protein;    gene expression;    immediate early genes;    live cell imaging;    spatio-temporal resolution;   

DOI  :  10.2144/btn-2019-0050

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown