Professor
UMR 7203 Laboratoire des Biomolécules (LBM)1,2,3
Sorbonne Université, Faculté des Sciences, Tour 33-23, 4 place Jussieu, 75005 Paris
1Sorbonne Université
2École Normale Supérieure – PSL Université
3CNRS
Email: arnaud.gautier@sorbonne-universite.fr
Website
Short bio
Graduated from the École Normale Supérieure of Lyon, I specialized in Chemical Biology in the labs of Prof. Kai Johnsson at the École Polytechnique Fédérale de Lausanne, and Prof. Jason Chin at the MRC Laboratory of Molecular Biology in Cambridge UK. After starting my independent career as Assistant Professor at the École Normale Supérieure in Paris, I joined Sorbonne University in Paris as Full Professor in Chemistry. My lab aims at developing Chemical Biology tools to push the frontiers of biological imaging.
Academic positions
2019 – now Professor of Chemistry at Sorbonne University (Paris, France), and Investigator in the Laboratory of Biomolecules (UMR 7203)
2010 – 2019 Assistant Professor of Chemistry at École Normale Supérieure – PSL University (Paris, France), and Investigator in the Laboratory PASTEUR (UMR 8640)
2009 – 2010 Postdoc, MRC Laboratory of Molecular Biology
2006 – 2009 Postdoc, École Polytechnique Fédérale de Lausanne
Education
2016 Habilitation (HDR) in Chemistry, École Normale Supérieure
2005 PhD in Chemistry, École Normale Supérieure de Lyon
2002 Master in Chemistry, École Normale Supérieure de Lyon /Université Lyon 1 Claude Bernard
Research interests
Cells and organisms are complex machines driven by a set of dynamic biological events tightly orchestrated in space and time. Our understanding of their inner workings is intricately related to our ability to observe how their constituents organize and interact. Despite spectacular developments in biological imaging and probe design, many biologically relevant molecules and processes remain invisible. Arnaud Gautier is interested in inventing new tools for observing biomolecules and dynamic biochemical events in live cells and tissues. These tools can allow biologists to address questions ranging from fundamental mechanisms to the causes of disease and the development of novel therapeutics.
Awards and distinctions
2018 Junior Member of the Institut Universitaire de France (IUF)
2017 CNRS Bronze Medal
2016 ERC Consolidator Laureate
2011 FEBS Distinguished Young Investigator Award
Publications
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. |
Reciprocal Regulation of Shh Trafficking and H2O2 Levels via a Noncanonical BOC-Rac1 Pathway Article de journal Marion Thauvin; Irène Amblard; Christine Rampon; Aurélien Mourton; Isabelle Queguiner; Chenge Li; Arnaud Gautier; Alain Joliot; Michel Volovitch; Sophie Vriz Antioxidants (Basel, Switzerland), 11 (4), p. 718, 2022, ISSN: 2076-3921. @article{thauvin_reciprocal_2022, title = {Reciprocal Regulation of Shh Trafficking and H2O2 Levels via a Noncanonical BOC-Rac1 Pathway}, author = {Marion Thauvin and Ir\`{e}ne Amblard and Christine Rampon and Aur\'{e}lien Mourton and Isabelle Queguiner and Chenge Li and Arnaud Gautier and Alain Joliot and Michel Volovitch and Sophie Vriz}, doi = {10.3390/antiox11040718}, issn = {2076-3921}, year = {2022}, date = {2022-01-01}, journal = {Antioxidants (Basel, Switzerland)}, volume = {11}, number = {4}, pages = {718}, abstract = {Among molecules that bridge environment, cell metabolism, and cell signaling, hydrogen peroxide (H2O2) recently appeared as an emerging but central player. Its level depends on cell metabolism and environment and was recently shown to play key roles during embryogenesis, contrasting with its long-established role in disease progression. We decided to explore whether the secreted morphogen Sonic hedgehog (Shh), known to be essential in a variety of biological processes ranging from embryonic development to adult tissue homeostasis and cancers, was part of these interactions. Here, we report that H2O2 levels control key steps of Shh delivery in cell culture: increased levels reduce primary secretion, stimulate endocytosis and accelerate delivery to recipient cells; in addition, physiological in vivo modulation of H2O2 levels changes Shh distribution and tissue patterning. Moreover, a feedback loop exists in which Shh trafficking controls H2O2 synthesis via a non-canonical BOC-Rac1 pathway, leading to cytoneme growth. Our findings reveal that Shh directly impacts its own distribution, thus providing a molecular explanation for the robustness of morphogenesis to both environmental insults and individual variability.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Among molecules that bridge environment, cell metabolism, and cell signaling, hydrogen peroxide (H2O2) recently appeared as an emerging but central player. Its level depends on cell metabolism and environment and was recently shown to play key roles during embryogenesis, contrasting with its long-established role in disease progression. We decided to explore whether the secreted morphogen Sonic hedgehog (Shh), known to be essential in a variety of biological processes ranging from embryonic development to adult tissue homeostasis and cancers, was part of these interactions. Here, we report that H2O2 levels control key steps of Shh delivery in cell culture: increased levels reduce primary secretion, stimulate endocytosis and accelerate delivery to recipient cells; in addition, physiological in vivo modulation of H2O2 levels changes Shh distribution and tissue patterning. Moreover, a feedback loop exists in which Shh trafficking controls H2O2 synthesis via a non-canonical BOC-Rac1 pathway, leading to cytoneme growth. Our findings reveal that Shh directly impacts its own distribution, thus providing a molecular explanation for the robustness of morphogenesis to both environmental insults and individual variability. |
2021 |
Orthogonal fluorescent chemogenetic reporters for multicolor imaging Article de journal Alison G Tebo; Benjamien Moeyaert; Marion Thauvin; Irene Carlon-Andres; Dorothea Böken; Michel Volovitch; Sergi Padilla-Parra; Peter Dedecker; Sophie Vriz; Arnaud Gautier Nature Chemical Biology, 17 (1), p. 30–38, 2021, ISSN: 1552-4469. @article{tebo_orthogonal_2021, title = {Orthogonal fluorescent chemogenetic reporters for multicolor imaging}, author = {Alison G Tebo and Benjamien Moeyaert and Marion Thauvin and Irene Carlon-Andres and Dorothea B\"{o}ken and Michel Volovitch and Sergi Padilla-Parra and Peter Dedecker and Sophie Vriz and Arnaud Gautier}, doi = {10.1038/s41589-020-0611-0}, issn = {1552-4469}, year = {2021}, date = {2021-01-01}, journal = {Nature Chemical Biology}, volume = {17}, number = {1}, pages = {30--38}, abstract = {Spectrally separated fluorophores allow the observation of multiple targets simultaneously inside living cells, leading to a deeper understanding of the molecular interplay that regulates cell function and fate. Chemogenetic systems combining a tag and a synthetic fluorophore provide certain advantages over fluorescent proteins since there is no requirement for chromophore maturation. Here, we present the engineering of a set of spectrally orthogonal fluorogen-activating tags based on the fluorescence-activating and absorption shifting tag (FAST) that are compatible with two-color, live-cell imaging. The resulting tags, greenFAST and redFAST, demonstrate orthogonality not only in their fluorogen recognition capabilities, but also in their one- and two-photon absorption profiles. This pair of orthogonal tags allowed the creation of a two-color cell cycle sensor capable of detecting very short, early cell cycles in zebrafish development and the development of split complementation systems capable of detecting multiple protein-protein interactions by live-cell fluorescence microscopy.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Spectrally separated fluorophores allow the observation of multiple targets simultaneously inside living cells, leading to a deeper understanding of the molecular interplay that regulates cell function and fate. Chemogenetic systems combining a tag and a synthetic fluorophore provide certain advantages over fluorescent proteins since there is no requirement for chromophore maturation. Here, we present the engineering of a set of spectrally orthogonal fluorogen-activating tags based on the fluorescence-activating and absorption shifting tag (FAST) that are compatible with two-color, live-cell imaging. The resulting tags, greenFAST and redFAST, demonstrate orthogonality not only in their fluorogen recognition capabilities, but also in their one- and two-photon absorption profiles. This pair of orthogonal tags allowed the creation of a two-color cell cycle sensor capable of detecting very short, early cell cycles in zebrafish development and the development of split complementation systems capable of detecting multiple protein-protein interactions by live-cell fluorescence microscopy. |
Arnaud Gautier; Ludovic Jullien; Chenge Li; Marie-Aude Plamont; Alison G Tebo; Marion Thauvin; Michel Volovitch; Sophie Vriz Springer US, New York, NY, 2021, ISBN: 978-1-07-161593-5. @book{gautier_versatile_2021, title = {Versatile On-Demand Fluorescent Labeling of Fusion Proteins Using Fluorescence-Activating and Absorption-Shifting Tag (FAST)}, author = {Arnaud Gautier and Ludovic Jullien and Chenge Li and Marie-Aude Plamont and Alison G Tebo and Marion Thauvin and Michel Volovitch and Sophie Vriz}, editor = {Eli Zamir}, url = {https://doi.org/10.1007/978-1-0716-1593-5_16}, doi = {10.1007/978-1-0716-1593-5_16}, isbn = {978-1-07-161593-5}, year = {2021}, date = {2021-01-01}, urldate = {2023-10-31}, booktitle = {Multiplexed Imaging: Methods and Protocols}, pages = {253--265}, publisher = {Springer US}, address = {New York, NY}, series = {Methods in Molecular Biology}, abstract = {Observing the localization, the concentration, and the distribution of proteins in cells or organisms is essential to understand theirs functions. General and versatile methods allowing multiplexed imaging of proteins under a large variety of experimental conditions are thus essential for deciphering the inner workings of cells and organisms. Here, we present a general method based on the non-covalent labeling of a small protein tag, named FAST (fluorescence-activating and absorption-shifting tag), with various fluorogenic ligands that light up upon labeling, which makes the simple, robust, and versatile on-demand labeling of fusion proteins in a wide range of experimental systems possible.}, keywords = {}, pubstate = {published}, tppubtype = {book} } Observing the localization, the concentration, and the distribution of proteins in cells or organisms is essential to understand theirs functions. General and versatile methods allowing multiplexed imaging of proteins under a large variety of experimental conditions are thus essential for deciphering the inner workings of cells and organisms. Here, we present a general method based on the non-covalent labeling of a small protein tag, named FAST (fluorescence-activating and absorption-shifting tag), with various fluorogenic ligands that light up upon labeling, which makes the simple, robust, and versatile on-demand labeling of fusion proteins in a wide range of experimental systems possible. |
2020 |
A Far-Red Emitting Fluorescent Chemogenetic Reporter for In Vivo Molecular Imaging Article de journal Chenge Li; Alison G Tebo; Marion Thauvin; Marie-Aude Plamont; Michel Volovitch; Xavier Morin; Sophie Vriz; Arnaud Gautier Angewandte Chemie International Edition, 59 (41), p. 17917–17923, 2020, ISSN: 1521-3773, (_eprint: https://onlinelibrary.wiley.com/doi/pdf/10.1002/anie.202006576). @article{li_far-red_2020, title = {A Far-Red Emitting Fluorescent Chemogenetic Reporter for In Vivo Molecular Imaging}, author = {Chenge Li and Alison G Tebo and Marion Thauvin and Marie-Aude Plamont and Michel Volovitch and Xavier Morin and Sophie Vriz and Arnaud Gautier}, url = {https://onlinelibrary.wiley.com/doi/abs/10.1002/anie.202006576}, doi = {10.1002/anie.202006576}, issn = {1521-3773}, year = {2020}, date = {2020-01-01}, urldate = {2023-10-31}, journal = {Angewandte Chemie International Edition}, volume = {59}, number = {41}, pages = {17917--17923}, abstract = {Far-red emitting fluorescent labels are highly desirable for spectral multiplexing and deep tissue imaging. Here, we describe the generation of frFAST (far-red Fluorescence Activating and absorption Shifting Tag), a 14-kDa monomeric protein that forms a bright far-red fluorescent assembly with (4-hydroxy-3-methoxy-phenyl)allylidene rhodanine (HPAR-3OM). As HPAR-3OM is essentially non-fluorescent in solution and in cells, frFAST can be imaged with high contrast in presence of free HPAR-3OM, which allowed the rapid and efficient imaging of frFAST fusions in live cells, zebrafish embryo/larvae, and chicken embryos. Beyond enabling the genetic encoding of far-red fluorescence, frFAST allowed the design of a far-red chemogenetic reporter of protein\textendashprotein interactions, demonstrating its great potential for the design of innovative far-red emitting biosensors.}, note = {_eprint: https://onlinelibrary.wiley.com/doi/pdf/10.1002/anie.202006576}, keywords = {}, pubstate = {published}, tppubtype = {article} } Far-red emitting fluorescent labels are highly desirable for spectral multiplexing and deep tissue imaging. Here, we describe the generation of frFAST (far-red Fluorescence Activating and absorption Shifting Tag), a 14-kDa monomeric protein that forms a bright far-red fluorescent assembly with (4-hydroxy-3-methoxy-phenyl)allylidene rhodanine (HPAR-3OM). As HPAR-3OM is essentially non-fluorescent in solution and in cells, frFAST can be imaged with high contrast in presence of free HPAR-3OM, which allowed the rapid and efficient imaging of frFAST fusions in live cells, zebrafish embryo/larvae, and chicken embryos. Beyond enabling the genetic encoding of far-red fluorescence, frFAST allowed the design of a far-red chemogenetic reporter of protein–protein interactions, demonstrating its great potential for the design of innovative far-red emitting biosensors. |
2019 |
A split fluorescent reporter with rapid and reversible complementation Article de journal Alison G Tebo; Arnaud Gautier Nature Communications, 10 (1), p. 2822, 2019, ISSN: 2041-1723. @article{Tebo2019, title = {A split fluorescent reporter with rapid and reversible complementation}, author = {Alison G Tebo and Arnaud Gautier}, url = {https://doi.org/10.1038/s41467-019-10855-0}, doi = {10.1038/s41467-019-10855-0}, issn = {2041-1723}, year = {2019}, date = {2019-01-01}, journal = {Nature Communications}, volume = {10}, number = {1}, pages = {2822}, abstract = {Interactions between proteins play an essential role in metabolic and signaling pathways, cellular processes and organismal systems. We report the development of splitFAST, a fluorescence complementation system for the visualization of transient protein-protein interactions in living cells. Engineered from the fluorogenic reporter FAST (Fluorescence-Activating and absorption-Shifting Tag), which specifically and reversibly binds fluorogenic hydroxybenzylidene rhodanine (HBR) analogs, splitFAST displays rapid and reversible complementation, allowing the real-time visualization of both the formation and the dissociation of a protein assembly.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Interactions between proteins play an essential role in metabolic and signaling pathways, cellular processes and organismal systems. We report the development of splitFAST, a fluorescence complementation system for the visualization of transient protein-protein interactions in living cells. Engineered from the fluorogenic reporter FAST (Fluorescence-Activating and absorption-Shifting Tag), which specifically and reversibly binds fluorogenic hydroxybenzylidene rhodanine (HBR) analogs, splitFAST displays rapid and reversible complementation, allowing the real-time visualization of both the formation and the dissociation of a protein assembly. |
Live Cell Super Resolution Imaging by Radial Fluctuations Using Fluorogen Binding Tags Article de journal Muthukumaran Venkatachalapathy; Vivek Belapurkar; Mini Jose; Arnaud Gautier; Deepak Nair Nanoscale, 11 (8), p. 3626-3632, 2019, ISSN: 2040-3364. @article{RN45, title = {Live Cell Super Resolution Imaging by Radial Fluctuations Using Fluorogen Binding Tags}, author = {Muthukumaran Venkatachalapathy and Vivek Belapurkar and Mini Jose and Arnaud Gautier and Deepak Nair}, doi = {10.1039/C8NR07809B}, issn = {2040-3364}, year = {2019}, date = {2019-01-01}, journal = {Nanoscale}, volume = {11}, number = {8}, pages = {3626-3632}, keywords = {}, pubstate = {published}, tppubtype = {article} } |
Single-Molecule Localization Microscopy with the Fluorescence-Activating and Absorption-Shifting Tag (FAST) System Article de journal Elizabeth M Smith; Arnaud Gautier; Elias M Puchner ACS Chemical Biology, 14 (6), p. 1115-1120, 2019, ISSN: 1554-8929. @article{Smith2019, title = {Single-Molecule Localization Microscopy with the Fluorescence-Activating and Absorption-Shifting Tag (FAST) System}, author = {Elizabeth M Smith and Arnaud Gautier and Elias M Puchner}, url = {https://doi.org/10.1021/acschembio.9b00149}, doi = {10.1021/acschembio.9b00149}, issn = {1554-8929}, year = {2019}, date = {2019-06-21}, journal = {ACS Chemical Biology}, volume = {14}, number = {6}, pages = {1115-1120}, publisher = {American Chemical Society}, abstract = {We develop and employ the Fluorescence-Activating and absorption-Shifting Tag (FAST) system for super-resolution (SR) imaging and single-molecule tracking based on single-molecule localizations. The fast off rate of fluorogen binding, combined with its spatially well-separated labeling of the densely expressed FAST fusion proteins, allowed single-molecule measurements to be performed in both living and fixed cells. The well-separated fluorescence localization density was achieved by either reversibly controlling the fluorogen concentration or by irreversibly photobleaching the FAST-fluorogen complex. The experimentally determined resolution of 28 nm allowed us to resolve Ensconsin-labeled microtubules and to track single molecules in mitochondria. Our results demonstrate that FAST is well-suited for single-molecule localization microscopy (SMLM). The small size and the availability of spectrally distinct fluorogens present unique advantages of the FAST system as a potential orthogonal labeling strategy that could be applied in conjunction with existing super-resolution dyes and photoactivatable proteins in versatile imaging applications.}, keywords = {}, pubstate = {published}, tppubtype = {article} } We develop and employ the Fluorescence-Activating and absorption-Shifting Tag (FAST) system for super-resolution (SR) imaging and single-molecule tracking based on single-molecule localizations. The fast off rate of fluorogen binding, combined with its spatially well-separated labeling of the densely expressed FAST fusion proteins, allowed single-molecule measurements to be performed in both living and fixed cells. The well-separated fluorescence localization density was achieved by either reversibly controlling the fluorogen concentration or by irreversibly photobleaching the FAST-fluorogen complex. The experimentally determined resolution of 28 nm allowed us to resolve Ensconsin-labeled microtubules and to track single molecules in mitochondria. Our results demonstrate that FAST is well-suited for single-molecule localization microscopy (SMLM). The small size and the availability of spectrally distinct fluorogens present unique advantages of the FAST system as a potential orthogonal labeling strategy that could be applied in conjunction with existing super-resolution dyes and photoactivatable proteins in versatile imaging applications. |
The Glowing Panoply of Fluorogen-based Markers for Advanced Bioimaging Book Chapter A Gautier The Glowing Panoply of Fluorogen-based Markers for Advanced Bioimaging, Chapitre 3, 2019. @inbook{Gautier:2019, title = {The Glowing Panoply of Fluorogen-based Markers for Advanced Bioimaging}, author = {A Gautier}, url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85056600039&doi=10.1039%2f9781788013284-00041&partnerID=40&md5=5d754dc76f6a9652835200ef4214a41a}, doi = {10.1039/9781788013284-00041}, year = {2019}, date = {2019-01-01}, booktitle = {The Glowing Panoply of Fluorogen-based Markers for Advanced Bioimaging}, chapter = {3}, series = {Comprehensive Series in Photochemical and Photobiological Sciences}, abstract = {The biological sciences nowadays rely extensively on imaging tools to decipher the complexity of living organisms. Fluorescence microscopy allows the study of biological processes with unprecedented temporal and spatial resolution. The revolution in fluorescence imaging has been the development of a large toolbox of fluorescent proteins able to reveal the abundance, position and dynamics of proteins. The fluorescence toolbox has recently been expanded with innovative reporters that permit proteins, and other biomolecules such as RNA, to be visualized in new ways. These innovative reporters are bipartite systems composed of a genetically encoded tag forming a fluorescent complex with a small organic fluorogenic chromophore (also called fluorogens). This chapter is a user-oriented presentation of some of the most mature fluorogen-based markers available to biologists. © 2019 European Society for Photobiology.}, keywords = {}, pubstate = {published}, tppubtype = {inbook} } The biological sciences nowadays rely extensively on imaging tools to decipher the complexity of living organisms. Fluorescence microscopy allows the study of biological processes with unprecedented temporal and spatial resolution. The revolution in fluorescence imaging has been the development of a large toolbox of fluorescent proteins able to reveal the abundance, position and dynamics of proteins. The fluorescence toolbox has recently been expanded with innovative reporters that permit proteins, and other biomolecules such as RNA, to be visualized in new ways. These innovative reporters are bipartite systems composed of a genetically encoded tag forming a fluorescent complex with a small organic fluorogenic chromophore (also called fluorogens). This chapter is a user-oriented presentation of some of the most mature fluorogen-based markers available to biologists. © 2019 European Society for Photobiology. |
2018 |
Circularly Permuted Fluorogenic Proteins for the Design of Modular Biosensors Article de journal A G Tebo; F M Pimenta; M Zoumpoulaki; C Kikuti; H Sirkia; M -A Plamont; A Houdusse; A Gautier ACS Chemical Biology, 13 (9), p. 2392–2397, 2018. @article{Tebo:2018, title = {Circularly Permuted Fluorogenic Proteins for the Design of Modular Biosensors}, author = {A G Tebo and F M Pimenta and M Zoumpoulaki and C Kikuti and H Sirkia and M -A Plamont and A Houdusse and A Gautier}, url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85052286419&doi=10.1021%2facschembio.8b00417&partnerID=40&md5=f7f8a46015d14cb7450f3d6d2b70a529}, doi = {10.1021/acschembio.8b00417}, year = {2018}, date = {2018-01-01}, journal = {ACS Chemical Biology}, volume = {13}, number = {9}, pages = {2392--2397}, abstract = {Fluorescent reporters are essential components for the design of optical biosensors that are able to image intracellular analytes in living cells. Herein, we describe the development of circularly permuted variants of Fluorescence-Activating and absorption-Shifting Tag (FAST) and demonstrate their potential as reporting module in biosensors. Circularly permutated FAST (cpFAST) variants allow one to condition the binding and activation of a fluorogenic ligand (and thus fluorescence) to analyte recognition by coupling them with analyte-binding domains. We demonstrated their use for biosensor design by generating multicolor plug-and-play fluorogenic biosensors for imaging the intracellular levels of Ca2+ in living mammalian cells in real time. © 2018 American Chemical Society.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Fluorescent reporters are essential components for the design of optical biosensors that are able to image intracellular analytes in living cells. Herein, we describe the development of circularly permuted variants of Fluorescence-Activating and absorption-Shifting Tag (FAST) and demonstrate their potential as reporting module in biosensors. Circularly permutated FAST (cpFAST) variants allow one to condition the binding and activation of a fluorogenic ligand (and thus fluorescence) to analyte recognition by coupling them with analyte-binding domains. We demonstrated their use for biosensor design by generating multicolor plug-and-play fluorogenic biosensors for imaging the intracellular levels of Ca2+ in living mammalian cells in real time. © 2018 American Chemical Society. |
Fluorogenic Probing of Membrane Protein Trafficking Article de journal C Li; A Mourton; M -A Plamont; V Rodrigues; I Aujard; M Volovitch; T Le Saux; F Perez; S Vriz; L Jullien; A Joliot; A Gautier Bioconjugate Chemistry, 29 (6), p. 1823–1828, 2018. @article{Li:2018a, title = {Fluorogenic Probing of Membrane Protein Trafficking}, author = {C Li and A Mourton and M -A Plamont and V Rodrigues and I Aujard and M Volovitch and T Le Saux and F Perez and S Vriz and L Jullien and A Joliot and A Gautier}, url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85047641063&doi=10.1021%2facs.bioconjchem.8b00180&partnerID=40&md5=bcbb74499b742fa8b0762b8c52ebf5dd}, doi = {10.1021/acs.bioconjchem.8b00180}, year = {2018}, date = {2018-01-01}, journal = {Bioconjugate Chemistry}, volume = {29}, number = {6}, pages = {1823--1828}, abstract = {Methods to differentially label cell-surface and intracellular membrane proteins are indispensable for understanding their function and the regulation of their trafficking. We present an efficient strategy for the rapid and selective fluorescent labeling of membrane proteins based on the chemical-genetic fluorescent marker FAST (fluorescence-activating and absorption-shifting tag). Cell-surface FAST-tagged proteins could be selectively and rapidly labeled using fluorogenic membrane-impermeant 4-hydroxybenzylidene rhodanine (HBR) analogs. This approach allows the study of protein trafficking at the plasma membrane with various fluorometric techniques, and opens exciting prospects for the high-throughput screening of small molecules able to restore disease-related trafficking defects. © 2018 American Chemical Society.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Methods to differentially label cell-surface and intracellular membrane proteins are indispensable for understanding their function and the regulation of their trafficking. We present an efficient strategy for the rapid and selective fluorescent labeling of membrane proteins based on the chemical-genetic fluorescent marker FAST (fluorescence-activating and absorption-shifting tag). Cell-surface FAST-tagged proteins could be selectively and rapidly labeled using fluorogenic membrane-impermeant 4-hydroxybenzylidene rhodanine (HBR) analogs. This approach allows the study of protein trafficking at the plasma membrane with various fluorometric techniques, and opens exciting prospects for the high-throughput screening of small molecules able to restore disease-related trafficking defects. © 2018 American Chemical Society. |
Fluorogenic Protein-Based Strategies for Detection, Actuation, and Sensing Article de journal A Gautier; A G Tebo BioEssays, 40 (10), 2018. @article{Gautier:2018, title = {Fluorogenic Protein-Based Strategies for Detection, Actuation, and Sensing}, author = {A Gautier and A G Tebo}, url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85053180336&doi=10.1002%2fbies.201800118&partnerID=40&md5=92409e91810fa2bf17d3eef357bc1bc9}, doi = {10.1002/bies.201800118}, year = {2018}, date = {2018-01-01}, journal = {BioEssays}, volume = {40}, number = {10}, abstract = {Fluorescence imaging has become an indispensable tool in cell and molecular biology. GFP-like fluorescent proteins have revolutionized fluorescence microscopy, giving experimenters exquisite control over the localization and specificity of tagged constructs. However, these systems present certain drawbacks and as such, alternative systems based on a fluorogenic interaction between a chromophore and a protein have been developed. While these systems are initially designed as fluorescent labels, they also present new opportunities for the development of novel labeling and detection strategies. This review focuses on new labeling protocols, actuation methods, and biosensors based on fluorogenic protein systems. © 2018 WILEY Periodicals, Inc.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Fluorescence imaging has become an indispensable tool in cell and molecular biology. GFP-like fluorescent proteins have revolutionized fluorescence microscopy, giving experimenters exquisite control over the localization and specificity of tagged constructs. However, these systems present certain drawbacks and as such, alternative systems based on a fluorogenic interaction between a chromophore and a protein have been developed. While these systems are initially designed as fluorescent labels, they also present new opportunities for the development of novel labeling and detection strategies. This review focuses on new labeling protocols, actuation methods, and biosensors based on fluorogenic protein systems. © 2018 WILEY Periodicals, Inc. |
Improved Chemical-Genetic Fluorescent Markers for Live Cell Microscopy Article de journal A G Tebo; F M Pimenta; Y Zhang; A Gautier Biochemistry, 57 (39), p. 5648–5653, 2018. @article{Tebo:2018a, title = {Improved Chemical-Genetic Fluorescent Markers for Live Cell Microscopy}, author = {A G Tebo and F M Pimenta and Y Zhang and A Gautier}, url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85053681817&doi=10.1021%2facs.biochem.8b00649&partnerID=40&md5=550c3f9cf88557339074b67771afd7d7}, doi = {10.1021/acs.biochem.8b00649}, year = {2018}, date = {2018-01-01}, journal = {Biochemistry}, volume = {57}, number = {39}, pages = {5648--5653}, abstract = {Inducible chemical-genetic fluorescent markers are promising tools for live cell imaging requiring high spatiotemporal resolution and low background fluorescence. The fluorescence-activating and absorption shifting tag (FAST) was recently developed to form fluorescent molecular complexes with a family of small, synthetic fluorogenic chromophores (so-called fluorogens). Here, we use rational design to modify the binding pocket of the protein and screen for improved fluorescence performances with four different fluorogens. The introduction of a single mutation results in improvements in both quantum yield and dissociation constant with nearly all fluorogens tested. Our improved FAST (iFAST) allowed the generation of a tandem iFAST (td-iFAST) that forms green and red fluorescent reporters 1.6-fold and 2-fold brighter than EGFP and mCherry, respectively, while having a comparable size. © Copyright 2018 American Chemical Society.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Inducible chemical-genetic fluorescent markers are promising tools for live cell imaging requiring high spatiotemporal resolution and low background fluorescence. The fluorescence-activating and absorption shifting tag (FAST) was recently developed to form fluorescent molecular complexes with a family of small, synthetic fluorogenic chromophores (so-called fluorogens). Here, we use rational design to modify the binding pocket of the protein and screen for improved fluorescence performances with four different fluorogens. The introduction of a single mutation results in improvements in both quantum yield and dissociation constant with nearly all fluorogens tested. Our improved FAST (iFAST) allowed the generation of a tandem iFAST (td-iFAST) that forms green and red fluorescent reporters 1.6-fold and 2-fold brighter than EGFP and mCherry, respectively, while having a comparable size. © Copyright 2018 American Chemical Society. |
Macroscale fluorescence imaging against autofluorescence under ambient light Article de journal R Zhang; R Chouket; M -A Plamont; Z Kelemen; A Espagne; A G Tebo; A Gautier; L Gissot; J -D Faure; L Jullien; V Croquette; T Le Saux Light: Science and Applications, 7 (1), 2018. @article{Zhang:2018c, title = {Macroscale fluorescence imaging against autofluorescence under ambient light}, author = {R Zhang and R Chouket and M -A Plamont and Z Kelemen and A Espagne and A G Tebo and A Gautier and L Gissot and J -D Faure and L Jullien and V Croquette and T Le Saux}, url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85058077105&doi=10.1038%2fs41377-018-0098-6&partnerID=40&md5=958a21f219ba413e687d1860d21c2767}, doi = {10.1038/s41377-018-0098-6}, year = {2018}, date = {2018-01-01}, journal = {Light: Science and Applications}, volume = {7}, number = {1}, abstract = {Macroscale fluorescence imaging is increasingly used to observe biological samples. However, it may suffer from spectral interferences that originate from ambient light or autofluorescence of the sample or its support. In this manuscript, we built a simple and inexpensive fluorescence macroscope, which has been used to evaluate the performance of Speed OPIOM (Out of Phase Imaging after Optical Modulation), which is a reference-free dynamic contrast protocol, to selectively image reversibly photoswitchable fluorophores as labels against detrimental autofluorescence and ambient light. By tuning the intensity and radial frequency of the modulated illumination to the Speed OPIOM resonance and adopting a phase-sensitive detection scheme that ensures noise rejection, we enhanced the sensitivity and the signal-to-noise ratio for fluorescence detection in blot assays by factors of 50 and 10, respectively, over direct fluorescence observation under constant illumination. Then, we overcame the strong autofluorescence of growth media that are currently used in microbiology and realized multiplexed fluorescence observation of colonies of spectrally similar fluorescent bacteria with a unique configuration of excitation and emission wavelengths. Finally, we easily discriminated fluorescent labels from the autofluorescent and reflective background in labeled leaves, even under the interference of incident light at intensities that are comparable to sunlight. The proposed approach is expected to find multiple applications, from biological assays to outdoor observations, in fluorescence macroimaging. © 2018, The Author(s).}, keywords = {}, pubstate = {published}, tppubtype = {article} } Macroscale fluorescence imaging is increasingly used to observe biological samples. However, it may suffer from spectral interferences that originate from ambient light or autofluorescence of the sample or its support. In this manuscript, we built a simple and inexpensive fluorescence macroscope, which has been used to evaluate the performance of Speed OPIOM (Out of Phase Imaging after Optical Modulation), which is a reference-free dynamic contrast protocol, to selectively image reversibly photoswitchable fluorophores as labels against detrimental autofluorescence and ambient light. By tuning the intensity and radial frequency of the modulated illumination to the Speed OPIOM resonance and adopting a phase-sensitive detection scheme that ensures noise rejection, we enhanced the sensitivity and the signal-to-noise ratio for fluorescence detection in blot assays by factors of 50 and 10, respectively, over direct fluorescence observation under constant illumination. Then, we overcame the strong autofluorescence of growth media that are currently used in microbiology and realized multiplexed fluorescence observation of colonies of spectrally similar fluorescent bacteria with a unique configuration of excitation and emission wavelengths. Finally, we easily discriminated fluorescent labels from the autofluorescent and reflective background in labeled leaves, even under the interference of incident light at intensities that are comparable to sunlight. The proposed approach is expected to find multiple applications, from biological assays to outdoor observations, in fluorescence macroimaging. © 2018, The Author(s). |
Spying on cells with chemical-genetic hybrids Article de journal A Gautier Actualite Chimique, (435), p. 31–35, 2018. @article{Gautier:2018b, title = {Spying on cells with chemical-genetic hybrids}, author = {A Gautier}, url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85059310973&partnerID=40&md5=89fdbcf594a9f3d4fe3ad8b1507ccef9}, year = {2018}, date = {2018-01-01}, journal = {Actualite Chimique}, number = {435}, pages = {31--35}, abstract = {Cells and organisms are complex machines regulated by a set of dynamic events orchestrated in space and time. Our understanding of their inner workings is intimately linked to our ability to observe how their constituents organize and interact. Optical microscopy allows to observe living systems at submicrometric scale. The development of high-performance fluorescent markers makes possible nowadays to monitor the dynamics of biomolecules with an unprecedented spatial and temporal resolution. This article presents how chemistry and biology can team up to develop next-generation markers that push the boundaries of biological imaging. © 2018 Societe Francaise de Chimie. All rights reserved.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Cells and organisms are complex machines regulated by a set of dynamic events orchestrated in space and time. Our understanding of their inner workings is intimately linked to our ability to observe how their constituents organize and interact. Optical microscopy allows to observe living systems at submicrometric scale. The development of high-performance fluorescent markers makes possible nowadays to monitor the dynamics of biomolecules with an unprecedented spatial and temporal resolution. This article presents how chemistry and biology can team up to develop next-generation markers that push the boundaries of biological imaging. © 2018 Societe Francaise de Chimie. All rights reserved. |