Practical and reliable FRET/FLIM pair of fluorescent proteins

Dmitry Shcherbo, Ekaterina A. Souslova, Joachim Goedhart, Tatyana V. Chepurnykh, Anna Gaintzeva, Irina I. Shemiakina, Theodorus W.J. Gadella, Sergey Lukyanov, Dmitriy M. Chudakov

Research output: Contribution to journalArticlepeer-review

78 Citations (Scopus)

Abstract

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.

Original languageEnglish
Article number24
JournalBMC Biotechnology
Volume9
DOIs
Publication statusPublished - 25 Mar 2009
Externally publishedYes

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