Practical and reliable FRET-FLIM pair of fluorescent proteinsReport as inadecuate




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BMC Biotechnology

, 9:24

First Online: 25 March 2009Received: 06 October 2008Accepted: 25 March 2009

Abstract

BackgroundIn 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.

ResultsHere 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.

ConclusionThe 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.

Electronic supplementary materialThe online version of this article doi:10.1186-1472-6750-9-24 contains supplementary material, which is available to authorized users.

Dmitry Shcherbo, Ekaterina A Souslova contributed equally to this work.

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Author: Dmitry Shcherbo - Ekaterina A Souslova - Joachim Goedhart - Tatyana V Chepurnykh - Anna Gaintzeva - Irina I Shemiakina -

Source: https://link.springer.com/article/10.1186/1472-6750-9-24



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