Practical and reliable FRET/FLIM pair of fluorescent proteins
<p>Abstract</p> <p>Background</p> <p>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...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2009-03-01
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| Series: | BMC Biotechnology |
| Online Access: | http://www.biomedcentral.com/1472-6750/9/24 |
| _version_ | 1818580270595440640 |
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| author | Shemiakina Irina I Gadella Theodorus WJ Gaintzeva Anna Chepurnykh Tatyana V Goedhart Joachim Souslova Ekaterina A Shcherbo Dmitry Lukyanov Sergey Chudakov Dmitriy M |
| author_facet | Shemiakina Irina I Gadella Theodorus WJ Gaintzeva Anna Chepurnykh Tatyana V Goedhart Joachim Souslova Ekaterina A Shcherbo Dmitry Lukyanov Sergey Chudakov Dmitriy M |
| author_sort | Shemiakina Irina I |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>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.</p> <p>Results</p> <p>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).</p> <p>Conclusion</p> <p>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.</p> |
| first_indexed | 2024-12-16T07:14:56Z |
| format | Article |
| id | doaj.art-5b7dfe47f058475bbfff1ce13f98f833 |
| institution | Directory Open Access Journal |
| issn | 1472-6750 |
| language | English |
| last_indexed | 2024-12-16T07:14:56Z |
| publishDate | 2009-03-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biotechnology |
| spelling | doaj.art-5b7dfe47f058475bbfff1ce13f98f8332022-12-21T22:39:47ZengBMCBMC Biotechnology1472-67502009-03-01912410.1186/1472-6750-9-24Practical and reliable FRET/FLIM pair of fluorescent proteinsShemiakina Irina IGadella Theodorus WJGaintzeva AnnaChepurnykh Tatyana VGoedhart JoachimSouslova Ekaterina AShcherbo DmitryLukyanov SergeyChudakov Dmitriy M<p>Abstract</p> <p>Background</p> <p>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.</p> <p>Results</p> <p>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).</p> <p>Conclusion</p> <p>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.</p>http://www.biomedcentral.com/1472-6750/9/24 |
| spellingShingle | Shemiakina Irina I Gadella Theodorus WJ Gaintzeva Anna Chepurnykh Tatyana V Goedhart Joachim Souslova Ekaterina A Shcherbo Dmitry Lukyanov Sergey Chudakov Dmitriy M Practical and reliable FRET/FLIM pair of fluorescent proteins BMC Biotechnology |
| title | Practical and reliable FRET/FLIM pair of fluorescent proteins |
| title_full | Practical and reliable FRET/FLIM pair of fluorescent proteins |
| title_fullStr | Practical and reliable FRET/FLIM pair of fluorescent proteins |
| title_full_unstemmed | Practical and reliable FRET/FLIM pair of fluorescent proteins |
| title_short | Practical and reliable FRET/FLIM pair of fluorescent proteins |
| title_sort | practical and reliable fret flim pair of fluorescent proteins |
| url | http://www.biomedcentral.com/1472-6750/9/24 |
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