期刊
BMC BIOTECHNOLOGY
卷 9, 期 -, 页码 -出版社
BMC
DOI: 10.1186/1472-6750-9-24
关键词
-
资金
- Howard Hughes Medical Institute [55005618]
- Molecular and Cell Biology Program RAS
- Rosnauka [02.512.11.2216]
- State Support of the Leading Scientific Schools [NS-2395.2008.4]
- President of Russian Federation [MK-6119.2008.4]
- [07-04-12189-ofi]
Background: In spite of a great number of monomeric fluorescent proteins developed in the recent years, the reported fluorescent protein-based FRET pairs are still characterized by a number of disadvantageous features, complicating their use as reporters in cell biology and for high-throughput cell-based screenings. Results: Here we screened some of the recently developed monomeric protein pairs to find the optimal combination, which would provide high dynamic range FRET changes, along with high pH- and photo-stability, fast maturation and bright fluorescence, and reliable detection in any fluorescent imaging system. Among generated FRET pairs, we have selected TagGFP-TagRFP, combining all the mentioned desirable characteristics. On the basis of this highly efficient FRET pair, we have generated a bright, high contrast, pH-and photo-stable apoptosis reporter, named CaspeR3 (Caspase 3 Reporter). Conclusion: The combined advantages suggest that the TagGFP-TagRFP is one of the most efficient green/red couples available to date for FRET/FLIM analyses to monitor interaction of proteins of interest in living cells and to generate FRET-based sensors for various applications. CaspeR3 provides reliable detection of apoptosis, and should become a popular tool both for cell biology studies and high throughput screening assays.
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