2024
|
A Fluorogenic Chemogenetic pH Sensor for Imaging Protein Exocytosis Article de journal Justine Coïs; Marie-Laure Niepon; Manon Wittwer; Hessam Sepasi Tehrani; Philippe Bun; Jean-Maurice Mallet; Vincent Vialou; Blaise Dumat ACS Sensors, 2024, ISSN: 2379-3694. @article{Cois2024b,
title = {A Fluorogenic Chemogenetic pH Sensor for Imaging Protein Exocytosis},
author = {Justine Co{\"{i}}s and Marie-Laure Niepon and Manon Wittwer and Hessam {Sepasi Tehrani} and Philippe Bun and Jean-Maurice Mallet and Vincent Vialou and Blaise Dumat},
url = {https://pubs.acs.org/doi/10.1021/acssensors.4c01057},
doi = {10.1021/acssensors.4c01057},
issn = {2379-3694},
year = {2024},
date = {2024-08-01},
journal = {ACS Sensors},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
|
Design of Bright Chemogenetic Reporters Based on the Combined Engineering of Fluorogenic Molecular Rotors and of the HaloTag Protein Article de journal Justine Coïs; Sylvestre P J T Bachollet; Louis Sanchez; Nicolas Pietrancosta; Vincent Vialou; Jean Maurice Mallet; Blaise Dumat Chemistry - A European Journal, 30 (32), p. e202400641, 2024, ISSN: 15213765. @article{Cois2024b,
title = {Design of Bright Chemogenetic Reporters Based on the Combined Engineering of Fluorogenic Molecular Rotors and of the HaloTag Protein},
author = {Justine Co{\"{i}}s and Sylvestre P J T Bachollet and Louis Sanchez and Nicolas Pietrancosta and Vincent Vialou and Jean Maurice Mallet and Blaise Dumat},
url = {https://chemistry-europe.onlinelibrary.wiley.com/doi/10.1002/chem.202400641},
doi = {10.1002/chem.202400641},
issn = {15213765},
year = {2024},
date = {2024-06-01},
journal = {Chemistry - A European Journal},
volume = {30},
number = {32},
pages = {e202400641},
abstract = {The combination of fluorogenic probes (fluorogens) and self-labeling protein tags represent a promising tool for imaging biological processes with high specificity but it requires the adequation between the fluorogen and its target to ensure a good activation of its fluorescence. In this work, we report a strategy to develop molecular rotors that specifically target HaloTag with a strong enhancement of their fluorescence. The divergent design facilitates the diversification of the structures to tune the photophysical and cellular properties. Four bright fluorogens with emissions ranging from green to red were identified and applied in wash-free live cell imaging experiments with good contrast and selectivity. A HaloTag mutant adapted from previous literature reports was also tested and shown to further improve the brightness and reaction rate of the most promising fluorogen of the series both in vitro and in cells. This work opens new possibilities to develop bright chemogenetic reporters with diverse photophysical and biological properties by exploring a potentially large chemical space of simple dipolar fluorophores in combination with protein engineering.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The combination of fluorogenic probes (fluorogens) and self-labeling protein tags represent a promising tool for imaging biological processes with high specificity but it requires the adequation between the fluorogen and its target to ensure a good activation of its fluorescence. In this work, we report a strategy to develop molecular rotors that specifically target HaloTag with a strong enhancement of their fluorescence. The divergent design facilitates the diversification of the structures to tune the photophysical and cellular properties. Four bright fluorogens with emissions ranging from green to red were identified and applied in wash-free live cell imaging experiments with good contrast and selectivity. A HaloTag mutant adapted from previous literature reports was also tested and shown to further improve the brightness and reaction rate of the most promising fluorogen of the series both in vitro and in cells. This work opens new possibilities to develop bright chemogenetic reporters with diverse photophysical and biological properties by exploring a potentially large chemical space of simple dipolar fluorophores in combination with protein engineering. |
FRET-Sensing of Multivalent Protein Binding at the Interface of Biomimetic Microparticles Functionalized with Fluorescent Glycolipids Article de journal Sophie Michelis; Chiara Pompili; Florence Niedergang; Jacques Fattaccioli; Blaise Dumat; Jean-Maurice Mallet ACS Applied Materials & Interfaces, 16 (8), p. 9669–9679, 2024, ISSN: 1944-8244. @article{Michelis2024,
title = {FRET-Sensing of Multivalent Protein Binding at the Interface of Biomimetic Microparticles Functionalized with Fluorescent Glycolipids},
author = {Sophie Michelis and Chiara Pompili and Florence Niedergang and Jacques Fattaccioli and Blaise Dumat and Jean-Maurice Mallet},
url = {https://pubs.acs.org/doi/10.1021/acsami.3c15067},
doi = {10.1021/acsami.3c15067},
issn = {1944-8244},
year = {2024},
date = {2024-02-01},
journal = {ACS Applied Materials & Interfaces},
volume = {16},
number = {8},
pages = {9669--9679},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
|
2022
|
An expanded palette of fluorogenic HaloTag probes with enhanced contrast for targeted cellular imaging Article de journal Sylvestre P J T Bachollet; Yuriy Shpinov; Fanny Broch; Hela Benaissa; Arnaud Gautier; Nicolas Pietrancosta; Jean-Maurice Mallet; Blaise Dumat Organic & Biomolecular Chemistry, 20 (17), p. 3619 - 3628, 2022, ISSN: 1477-0520. @article{Bachollet2022,
title = {An expanded palette of fluorogenic HaloTag probes with enhanced contrast for targeted cellular imaging},
author = {Sylvestre P J T Bachollet and Yuriy Shpinov and Fanny Broch and Hela Benaissa and Arnaud Gautier and Nicolas Pietrancosta and Jean-Maurice Mallet and Blaise Dumat},
url = {http://xlink.rsc.org/?DOI=D1OB02394B},
doi = {10.1039/D1OB02394B},
issn = {1477-0520},
year = {2022},
date = {2022-01-01},
journal = {Organic & Biomolecular Chemistry},
volume = {20},
number = {17},
pages = {3619 - 3628},
publisher = {Royal Society of Chemistry},
abstract = {A palette of fluorogenic molecular rotor probes with emissions from green to NIR was developed for wash-free and multicolor imaging of genetically-encoded HaloTag fusion proteins.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
A palette of fluorogenic molecular rotor probes with emissions from green to NIR was developed for wash-free and multicolor imaging of genetically-encoded HaloTag fusion proteins. |
Fluorogenic and Genetic Targeting of a Red-Emitting Molecular Calcium Indicator Article de journal Sylvestre P J T Bachollet; Nicolas Pietrancosta; Jean-Maurice Mallet; Blaise Dumat Chemical Communications, 2022. @article{Bachollet2022b,
title = {Fluorogenic and Genetic Targeting of a Red-Emitting Molecular Calcium Indicator},
author = {Sylvestre P J T Bachollet and Nicolas Pietrancosta and Jean-Maurice Mallet and Blaise Dumat},
doi = {10.1039/D2CC01792J},
year = {2022},
date = {2022-01-01},
journal = {Chemical Communications},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
|
2020
|
Fluorogenic Protein Probes with Red and Near‐Infrared Emission for Genetically Targeted Imaging** Article de journal Sylvestre P J T Bachollet; Cyril Addi; Nicolas Pietrancosta; Jean-Maurice Mallet; Blaise Dumat Chemistry – A European Journal, 26 (63), p. 14467–14473, 2020, ISSN: 0947-6539. @article{Bachollet2020,
title = {Fluorogenic Protein Probes with Red and Near‐Infrared Emission for Genetically Targeted Imaging**},
author = {Sylvestre P J T Bachollet and Cyril Addi and Nicolas Pietrancosta and Jean-Maurice Mallet and Blaise Dumat},
url = {https://onlinelibrary.wiley.com/doi/abs/10.1002/chem.202002911 https://onlinelibrary.wiley.com/doi/10.1002/chem.202002911},
doi = {10.1002/chem.202002911},
issn = {0947-6539},
year = {2020},
date = {2020-11-01},
journal = {Chemistry \textendash A European Journal},
volume = {26},
number = {63},
pages = {14467--14473},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
|
2019
|
Mannose-Coated Fluorescent Lipid Microparticles for Specific Cellular Targeting and Internalization via Glycoreceptor-Induced Phagocytosis Article de journal Blaise Dumat; Lorraine Montel; Léa Pinon; Pascal Matton; Laurent Cattiaux; Jacques Fattaccioli; Jean-Maurice Mallet ACS Applied Bio Materials, 2 , p. 5118-5126, 2019, ISSN: 2576-6422. @article{Dumat2019,
title = {Mannose-Coated Fluorescent Lipid Microparticles for Specific Cellular Targeting and Internalization via Glycoreceptor-Induced Phagocytosis},
author = {Blaise Dumat and Lorraine Montel and L\'{e}a Pinon and Pascal Matton and Laurent Cattiaux and Jacques Fattaccioli and Jean-Maurice Mallet},
url = {https://pubs.acs.org/doi/10.1021/acsabm.9b00793},
doi = {10.1021/acsabm.9b00793},
issn = {2576-6422},
year = {2019},
date = {2019-11-01},
journal = {ACS Applied Bio Materials},
volume = {2},
pages = {5118-5126},
publisher = {American Chemical Society},
abstract = {In this work we report on the development of mannose-coated fluorescent lipid microparticles to study the role of C-type lectin membrane receptor in phagocytosis. The micrometric droplets of Soybean oil in water emulsion were functionalized with a tailor-made fluorescent mannolipid. The amphiphilic ligand was built from a mannose unit, a lipid C11 spacer and a naphthalimide fluorophore. The droplets functionalization was monitored by fluorescence microscopy as well as the interaction with concanavalin A which was used as a model lectin in vitro. The use of a monovalent ligand on the surface of emulsion droplets yielded particles with an affinity approximately 40 times higher than that of free mannose. In cellulo, the coated droplets were shown to be specifically internalized by macrophages in a receptor-dependent phagocytic pathway. The naked droplets on the other hand display very little internalization due to their low immunogenicity. This work thus brings evidence that C-type lectin membrane receptors ...},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
In this work we report on the development of mannose-coated fluorescent lipid microparticles to study the role of C-type lectin membrane receptor in phagocytosis. The micrometric droplets of Soybean oil in water emulsion were functionalized with a tailor-made fluorescent mannolipid. The amphiphilic ligand was built from a mannose unit, a lipid C11 spacer and a naphthalimide fluorophore. The droplets functionalization was monitored by fluorescence microscopy as well as the interaction with concanavalin A which was used as a model lectin in vitro. The use of a monovalent ligand on the surface of emulsion droplets yielded particles with an affinity approximately 40 times higher than that of free mannose. In cellulo, the coated droplets were shown to be specifically internalized by macrophages in a receptor-dependent phagocytic pathway. The naked droplets on the other hand display very little internalization due to their low immunogenicity. This work thus brings evidence that C-type lectin membrane receptors ... |
2018
|
Synthesis, Oligonucleotide Incorporation and Fluorescence Properties in DNA of a Bicyclic Thymine Analogue Article de journal Christopher P Lawson; Anders F Füchtbauer; Moa S Wranne; Tristan Giraud; Thomas Floyd; Blaise Dumat; Nicolai K Andersen; Afaf H. El-Sagheer; Tom Brown; Henrik Gradén; Marcus L Wilhelmsson; Morten Grøtli Scientific Reports, 8 (1), p. 1-9, 2018, ISSN: 4159801831897. @article{Lawson:2018,
title = {Synthesis, Oligonucleotide Incorporation and Fluorescence Properties in DNA of a Bicyclic Thymine Analogue},
author = {Christopher P Lawson and Anders F F\"{u}chtbauer and Moa S Wranne and Tristan Giraud and Thomas Floyd and Blaise Dumat and Nicolai K Andersen and Afaf {H. El-Sagheer} and Tom Brown and Henrik Grad\'{e}n and Marcus L Wilhelmsson and Morten Gr\otli},
doi = {10.1038/s41598-018-31897-2},
issn = {4159801831897},
year = {2018},
date = {2018-01-01},
journal = {Scientific Reports},
volume = {8},
number = {1},
pages = {1-9},
abstract = {Fluorescent base analogues (FBAs) have emerged as a powerful class of molecular reporters of location and environment for nucleic acids. In our overall mission to develop bright and useful FBAs for all natural nucleobases, herein we describe the synthesis and thorough characterization of bicyclic thymidine (bT), both as a monomer and when incorporated into DNA. We have developed a robust synthetic route for the preparation of the bT DNA monomer and the corresponding protected phosphoramidite for solid-phase DNA synthesis. The bT deoxyribonucleoside has a brightness value of 790 M-1cm-1 in water, which is comparable or higher than most fluorescent thymine analogues reported. When incorporated into DNA, bT pairs selectively with adenine without perturbing the B-form structure, keeping the melting thermodynamics of the B-form duplex DNA virtually unchanged. As for most fluorescent base analogues, the emission of bT is reduced inside DNA (4.5- and 13-fold in single- and double-stranded DNA, respectively). Overall, these properties make bT an interesting thymine analogue for studying DNA and an excellent starting point for the development of brighter bT derivatives.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Fluorescent base analogues (FBAs) have emerged as a powerful class of molecular reporters of location and environment for nucleic acids. In our overall mission to develop bright and useful FBAs for all natural nucleobases, herein we describe the synthesis and thorough characterization of bicyclic thymidine (bT), both as a monomer and when incorporated into DNA. We have developed a robust synthetic route for the preparation of the bT DNA monomer and the corresponding protected phosphoramidite for solid-phase DNA synthesis. The bT deoxyribonucleoside has a brightness value of 790 M-1cm-1 in water, which is comparable or higher than most fluorescent thymine analogues reported. When incorporated into DNA, bT pairs selectively with adenine without perturbing the B-form structure, keeping the melting thermodynamics of the B-form duplex DNA virtually unchanged. As for most fluorescent base analogues, the emission of bT is reduced inside DNA (4.5- and 13-fold in single- and double-stranded DNA, respectively). Overall, these properties make bT an interesting thymine analogue for studying DNA and an excellent starting point for the development of brighter bT derivatives. |
2017
|
Toward Complete Sequence Flexibility of Nucleic Acid Base Analogue FRET Article de journal Moa S Wranne; Anders Foller Füchtbauer; Blaise Dumat; Mattias Bood; Afaf H El-Sagheer; Tom Brown; Henrik Gradén; Morten Grøtli; Marcus L Wilhelmsson Journal of the American Chemical Society, 139 (27), p. 9271-9280, 2017. @article{Wranne:2017,
title = {Toward Complete Sequence Flexibility of Nucleic Acid Base Analogue FRET},
author = {Moa S Wranne and Anders Foller F\"{u}chtbauer and Blaise Dumat and Mattias Bood and Afaf H {El-Sagheer} and Tom Brown and Henrik Grad\'{e}n and Morten Gr\otli and Marcus L Wilhelmsson},
doi = {10.1021/jacs.7b04517},
year = {2017},
date = {2017-01-01},
journal = {Journal of the American Chemical Society},
volume = {139},
number = {27},
pages = {9271-9280},
abstract = {F\"{o}rster resonance energy transfer (FRET) using fluorescent base analogues is a powerful means of obtaining high-resolution nucleic acid structure and dynamics information that favorably complements techniques such as NMR and X-ray crystallography. Here, we expand the base\textendashbase FRET repertoire with an adenine analogue FRET-pair. Phosphoramidite-protected quadracyclic 2$'$-deoxyadenosine analogues qAN1 (donor) and qAnitro (acceptor) were synthesized and incorporated into DNA by a generic, reliable, and high-yielding route, and both constitute excellent adenine analogues. The donor, qAN1, has quantum yields reaching 21% and 11% in single- and double-strands, respectively. To the best of our knowledge, this results in the highest average brightness of an adenine analogue inside DNA. Its potent emissive features overlap well with the absorption of qAnitro and thus enable accurate FRET-measurements over more than one turn of B-DNA. As we have shown previously for our cytosine analogue FRET-pair, FRET between qAN1...},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Förster resonance energy transfer (FRET) using fluorescent base analogues is a powerful means of obtaining high-resolution nucleic acid structure and dynamics information that favorably complements techniques such as NMR and X-ray crystallography. Here, we expand the base–base FRET repertoire with an adenine analogue FRET-pair. Phosphoramidite-protected quadracyclic 2$'$-deoxyadenosine analogues qAN1 (donor) and qAnitro (acceptor) were synthesized and incorporated into DNA by a generic, reliable, and high-yielding route, and both constitute excellent adenine analogues. The donor, qAN1, has quantum yields reaching 21% and 11% in single- and double-strands, respectively. To the best of our knowledge, this results in the highest average brightness of an adenine analogue inside DNA. Its potent emissive features overlap well with the absorption of qAnitro and thus enable accurate FRET-measurements over more than one turn of B-DNA. As we have shown previously for our cytosine analogue FRET-pair, FRET between qAN1... |
2016
|
Studying Z-DNA and B- to Z-DNA Transitions Using a Cytosine Analogue FRET-Pair Article de journal Blaise Dumat; Anders Foller Larsen; Marcus L Wilhelmsson Nucleic Acids Research, 44 (11), p. e101-e101, 2016. @article{Dumat:2016,
title = {Studying Z-DNA and B- to Z-DNA Transitions Using a Cytosine Analogue FRET-Pair},
author = {Blaise Dumat and Anders Foller Larsen and Marcus L Wilhelmsson},
doi = {10.1093/nar/gkw114},
year = {2016},
date = {2016-06-01},
journal = {Nucleic Acids Research},
volume = {44},
number = {11},
pages = {e101-e101},
abstract = {Herein, we report on the use of a tricyclic cytosine FRET pair, incorporated into DNA with different base pair separations, to study Z-DNA and B-Z DNA junc-tions. With its position inside the DNA structure, the FRET pair responds to a B-to Z-DNA transi-tion with a distinct change in FRET efficiency for each donor/acceptor configuration allowing reliable structural probing. Moreover, we show how fluores-cence spectroscopy and our cytosine analogues can be used to determine rate constants for the B-to Z-DNA transition mechanism. The modified cytosines have little influence on the transition and the FRET pair is thus an easily implemented and virtually non-perturbing fluorescence tool to study Z-DNA. This nucleobase analogue FRET pair represents a valu-able addition to the limited number of fluorescence methods available to study Z-DNA and we suggest it will facilitate, for example, deciphering the B-to Z-DNA transition mechanism and investigating the interaction of DNA with Z-DNA binding proteins.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Herein, we report on the use of a tricyclic cytosine FRET pair, incorporated into DNA with different base pair separations, to study Z-DNA and B-Z DNA junc-tions. With its position inside the DNA structure, the FRET pair responds to a B-to Z-DNA transi-tion with a distinct change in FRET efficiency for each donor/acceptor configuration allowing reliable structural probing. Moreover, we show how fluores-cence spectroscopy and our cytosine analogues can be used to determine rate constants for the B-to Z-DNA transition mechanism. The modified cytosines have little influence on the transition and the FRET pair is thus an easily implemented and virtually non-perturbing fluorescence tool to study Z-DNA. This nucleobase analogue FRET pair represents a valu-able addition to the limited number of fluorescence methods available to study Z-DNA and we suggest it will facilitate, for example, deciphering the B-to Z-DNA transition mechanism and investigating the interaction of DNA with Z-DNA binding proteins. |
