The Experts below are selected from a list of 47058 Experts worldwide ranked by ideXlab platform
Michael Ryckelynck - One of the best experts on this subject based on the ideXlab platform.
-
Structure and functional reselection of the Mango-III fluorogenic RNA aptamer
Nature Chemical Biology, 2019Co-Authors: Robert Trachman, Alexis Autour, Sunny Jeng, Amir Abdolahzadeh, Alessio Andreoni, Razvan Cojocaru, Ramil Garipov, Elena Dolgosheina, Jay Knutson, Michael RyckelynckAbstract:Several turn-on RNA aptamers that activate small molecule fluorophores have been selected in vitro. Among these, the ~30 nucleotide Mango-III is notable because it binds the thiazole orange derivative TO1-Biotin with high affinity and fluoresces brightly (quantum yield 0.55). Uniquely among related aptamers, Mango-III exhibits biphasic thermal melting, characteristic of molecules with tertiary structure. We report crystal structures of TO1-Biotin complexes of Mango-III, a structure-guided mutant Mango-III(A10U), and a functionally reselected mutant iMango-III. The structures reveal a globular architecture arising from an unprecedented pseudoknot-like connectivity between a G-quadruplex and an embedded non-canonical duplex. The fluorophore is restrained into a planar conformation by the G-quadruplex, a lone, long-range trans-Watson-Crick pair (whose A10U mutation increases quantum yield to 0.66), and a pyrimidine perpendicular to the nucleobase planes of those motifs. The improved iMango-III and Mango-III(A10U) fluoresce ~50% brighter than enhanced green fluorescent protein, making them suitable tags for live cell RNA visualization.
-
Structure and functional reselection of the Mango-III fluorogenic RNA aptamer.
Nature chemical biology, 2019Co-Authors: R.j. Trachoma, Alexis Autour, Amir Abdolahzadeh, Alessio Andreoni, Razvan Cojocaru, Ramil Garipov, Elena Dolgosheina, Sunny C. Y. Jeng, Jay R. Knutson, Michael RyckelynckAbstract:Several turn-on RNA aptamers that activate small-molecule fluorophores have been selected in vitro. Among these, the ~30 nucleotide Mango-III is notable because it binds the thiazole orange derivative TO1-Biotin with high affinity and fluoresces brightly (quantum yield 0.55). Uniquely among related aptamers, Mango-III exhibits biphasic thermal melting, characteristic of molecules with tertiary structure. We report crystal structures of TO1-Biotin complexes of Mango-III, a structure-guided mutant Mango-III(A10U), and a functionally reselected mutant iMango-III. The structures reveal a globular architecture arising from an unprecedented pseudoknot-like connectivity between a G-quadruplex and an embedded non-canonical duplex. The fluorophore is restrained into a planar conformation by the G-quadruplex, a lone, long-range trans Watson–Crick pair (whose A10U mutation increases quantum yield to 0.66), and a pyrimidine perpendicular to the nucleobase planes of those motifs. The improved iMango-III and Mango-III(A10U) fluoresce ~50% brighter than enhanced green fluorescent protein, making them suitable tags for live cell RNA visualization. Structural analysis of Mango-III in complex with fluorophores reveals a globular architecture and a planar conformation of fluorophores. Structure-guided mutation and functional reselection identified two mutants with improved fluorescence intensity.
-
Fluorogenic RNA Mango aptamers for imaging small non-coding RNAs in mammalian cells
Nature Communications, 2018Co-Authors: Alexis Autour, Sunny Jeng, Amir Abdolahzadeh, Michael Ryckelynck, Adam Cawte, Angela Galli, Shanker S. S. Panchapakesan, David Rueda, Peter J. UnrauAbstract:Despite having many key roles in cellular biology, directly imaging biologically important RNAs has been hindered by a lack of fluorescent tools equivalent to the fluorescent proteins available to study cellular proteins. Ideal RNA labelling systems must preserve biological function, have photophysical properties similar to existing fluorescent proteins, and be compatible with established live and fixed cell protein labelling strategies. Here, we report a microfluidics-based selection of three new high-affinity RNA Mango fluorogenic aptamers. Two of these are as bright or brighter than enhanced GFP when bound to TO1-Biotin. Furthermore, we show that the new Mangos can accurately image the subcellular localization of three small non-coding RNAs (5S, U6, and a box C/D scaRNA) in fixed and live mammalian cells. These new aptamers have many potential applications to study RNA function and dynamics both in vitro and in mammalian cells. Many aptamer-fluorophore complexes suffer from low quantum yield and low extinction coefficients limiting their usability. Here the authors isolate new Mango aptamers with improved fluorescent properties for both fixed and live-cell imaging by using competitive ligand binding microfluidic selection.
-
fluorogenic rna Mango aptamers for imaging small non coding rnas in mammalian cells
Nature Communications, 2018Co-Authors: Alexis Autour, Amir Abdolahzadeh, Michael Ryckelynck, Sunny C. Y. Jeng, Adam Cawte, Angela Galli, Shanker S. S. Panchapakesan, David Rueda, Peter J. UnrauAbstract:Despite having many key roles in cellular biology, directly imaging biologically important RNAs has been hindered by a lack of fluorescent tools equivalent to the fluorescent proteins available to study cellular proteins. Ideal RNA labelling systems must preserve biological function, have photophysical properties similar to existing fluorescent proteins, and be compatible with established live and fixed cell protein labelling strategies. Here, we report a microfluidics-based selection of three new high-affinity RNA Mango fluorogenic aptamers. Two of these are as bright or brighter than enhanced GFP when bound to TO1-Biotin. Furthermore, we show that the new Mangos can accurately image the subcellular localization of three small non-coding RNAs (5S, U6, and a box C/D scaRNA) in fixed and live mammalian cells. These new aptamers have many potential applications to study RNA function and dynamics both in vitro and in mammalian cells.
Alexis Autour - One of the best experts on this subject based on the ideXlab platform.
-
Structure and functional reselection of the Mango-III fluorogenic RNA aptamer
Nature Chemical Biology, 2019Co-Authors: Robert Trachman, Alexis Autour, Sunny Jeng, Amir Abdolahzadeh, Alessio Andreoni, Razvan Cojocaru, Ramil Garipov, Elena Dolgosheina, Jay Knutson, Michael RyckelynckAbstract:Several turn-on RNA aptamers that activate small molecule fluorophores have been selected in vitro. Among these, the ~30 nucleotide Mango-III is notable because it binds the thiazole orange derivative TO1-Biotin with high affinity and fluoresces brightly (quantum yield 0.55). Uniquely among related aptamers, Mango-III exhibits biphasic thermal melting, characteristic of molecules with tertiary structure. We report crystal structures of TO1-Biotin complexes of Mango-III, a structure-guided mutant Mango-III(A10U), and a functionally reselected mutant iMango-III. The structures reveal a globular architecture arising from an unprecedented pseudoknot-like connectivity between a G-quadruplex and an embedded non-canonical duplex. The fluorophore is restrained into a planar conformation by the G-quadruplex, a lone, long-range trans-Watson-Crick pair (whose A10U mutation increases quantum yield to 0.66), and a pyrimidine perpendicular to the nucleobase planes of those motifs. The improved iMango-III and Mango-III(A10U) fluoresce ~50% brighter than enhanced green fluorescent protein, making them suitable tags for live cell RNA visualization.
-
Structure and functional reselection of the Mango-III fluorogenic RNA aptamer.
Nature chemical biology, 2019Co-Authors: R.j. Trachoma, Alexis Autour, Amir Abdolahzadeh, Alessio Andreoni, Razvan Cojocaru, Ramil Garipov, Elena Dolgosheina, Sunny C. Y. Jeng, Jay R. Knutson, Michael RyckelynckAbstract:Several turn-on RNA aptamers that activate small-molecule fluorophores have been selected in vitro. Among these, the ~30 nucleotide Mango-III is notable because it binds the thiazole orange derivative TO1-Biotin with high affinity and fluoresces brightly (quantum yield 0.55). Uniquely among related aptamers, Mango-III exhibits biphasic thermal melting, characteristic of molecules with tertiary structure. We report crystal structures of TO1-Biotin complexes of Mango-III, a structure-guided mutant Mango-III(A10U), and a functionally reselected mutant iMango-III. The structures reveal a globular architecture arising from an unprecedented pseudoknot-like connectivity between a G-quadruplex and an embedded non-canonical duplex. The fluorophore is restrained into a planar conformation by the G-quadruplex, a lone, long-range trans Watson–Crick pair (whose A10U mutation increases quantum yield to 0.66), and a pyrimidine perpendicular to the nucleobase planes of those motifs. The improved iMango-III and Mango-III(A10U) fluoresce ~50% brighter than enhanced green fluorescent protein, making them suitable tags for live cell RNA visualization. Structural analysis of Mango-III in complex with fluorophores reveals a globular architecture and a planar conformation of fluorophores. Structure-guided mutation and functional reselection identified two mutants with improved fluorescence intensity.
-
Fluorogenic RNA Mango aptamers for imaging small non-coding RNAs in mammalian cells
Nature Communications, 2018Co-Authors: Alexis Autour, Sunny Jeng, Amir Abdolahzadeh, Michael Ryckelynck, Adam Cawte, Angela Galli, Shanker S. S. Panchapakesan, David Rueda, Peter J. UnrauAbstract:Despite having many key roles in cellular biology, directly imaging biologically important RNAs has been hindered by a lack of fluorescent tools equivalent to the fluorescent proteins available to study cellular proteins. Ideal RNA labelling systems must preserve biological function, have photophysical properties similar to existing fluorescent proteins, and be compatible with established live and fixed cell protein labelling strategies. Here, we report a microfluidics-based selection of three new high-affinity RNA Mango fluorogenic aptamers. Two of these are as bright or brighter than enhanced GFP when bound to TO1-Biotin. Furthermore, we show that the new Mangos can accurately image the subcellular localization of three small non-coding RNAs (5S, U6, and a box C/D scaRNA) in fixed and live mammalian cells. These new aptamers have many potential applications to study RNA function and dynamics both in vitro and in mammalian cells. Many aptamer-fluorophore complexes suffer from low quantum yield and low extinction coefficients limiting their usability. Here the authors isolate new Mango aptamers with improved fluorescent properties for both fixed and live-cell imaging by using competitive ligand binding microfluidic selection.
-
fluorogenic rna Mango aptamers for imaging small non coding rnas in mammalian cells
Nature Communications, 2018Co-Authors: Alexis Autour, Amir Abdolahzadeh, Michael Ryckelynck, Sunny C. Y. Jeng, Adam Cawte, Angela Galli, Shanker S. S. Panchapakesan, David Rueda, Peter J. UnrauAbstract:Despite having many key roles in cellular biology, directly imaging biologically important RNAs has been hindered by a lack of fluorescent tools equivalent to the fluorescent proteins available to study cellular proteins. Ideal RNA labelling systems must preserve biological function, have photophysical properties similar to existing fluorescent proteins, and be compatible with established live and fixed cell protein labelling strategies. Here, we report a microfluidics-based selection of three new high-affinity RNA Mango fluorogenic aptamers. Two of these are as bright or brighter than enhanced GFP when bound to TO1-Biotin. Furthermore, we show that the new Mangos can accurately image the subcellular localization of three small non-coding RNAs (5S, U6, and a box C/D scaRNA) in fixed and live mammalian cells. These new aptamers have many potential applications to study RNA function and dynamics both in vitro and in mammalian cells.
Peter J. Unrau - One of the best experts on this subject based on the ideXlab platform.
-
Fluorogenic RNA Mango aptamers for imaging small non-coding RNAs in mammalian cells
Nature Communications, 2018Co-Authors: Alexis Autour, Sunny Jeng, Amir Abdolahzadeh, Michael Ryckelynck, Adam Cawte, Angela Galli, Shanker S. S. Panchapakesan, David Rueda, Peter J. UnrauAbstract:Despite having many key roles in cellular biology, directly imaging biologically important RNAs has been hindered by a lack of fluorescent tools equivalent to the fluorescent proteins available to study cellular proteins. Ideal RNA labelling systems must preserve biological function, have photophysical properties similar to existing fluorescent proteins, and be compatible with established live and fixed cell protein labelling strategies. Here, we report a microfluidics-based selection of three new high-affinity RNA Mango fluorogenic aptamers. Two of these are as bright or brighter than enhanced GFP when bound to TO1-Biotin. Furthermore, we show that the new Mangos can accurately image the subcellular localization of three small non-coding RNAs (5S, U6, and a box C/D scaRNA) in fixed and live mammalian cells. These new aptamers have many potential applications to study RNA function and dynamics both in vitro and in mammalian cells. Many aptamer-fluorophore complexes suffer from low quantum yield and low extinction coefficients limiting their usability. Here the authors isolate new Mango aptamers with improved fluorescent properties for both fixed and live-cell imaging by using competitive ligand binding microfluidic selection.
-
fluorogenic rna Mango aptamers for imaging small non coding rnas in mammalian cells
Nature Communications, 2018Co-Authors: Alexis Autour, Amir Abdolahzadeh, Michael Ryckelynck, Sunny C. Y. Jeng, Adam Cawte, Angela Galli, Shanker S. S. Panchapakesan, David Rueda, Peter J. UnrauAbstract:Despite having many key roles in cellular biology, directly imaging biologically important RNAs has been hindered by a lack of fluorescent tools equivalent to the fluorescent proteins available to study cellular proteins. Ideal RNA labelling systems must preserve biological function, have photophysical properties similar to existing fluorescent proteins, and be compatible with established live and fixed cell protein labelling strategies. Here, we report a microfluidics-based selection of three new high-affinity RNA Mango fluorogenic aptamers. Two of these are as bright or brighter than enhanced GFP when bound to TO1-Biotin. Furthermore, we show that the new Mangos can accurately image the subcellular localization of three small non-coding RNAs (5S, U6, and a box C/D scaRNA) in fixed and live mammalian cells. These new aptamers have many potential applications to study RNA function and dynamics both in vitro and in mammalian cells.
Amir Abdolahzadeh - One of the best experts on this subject based on the ideXlab platform.
-
Structure and functional reselection of the Mango-III fluorogenic RNA aptamer
Nature Chemical Biology, 2019Co-Authors: Robert Trachman, Alexis Autour, Sunny Jeng, Amir Abdolahzadeh, Alessio Andreoni, Razvan Cojocaru, Ramil Garipov, Elena Dolgosheina, Jay Knutson, Michael RyckelynckAbstract:Several turn-on RNA aptamers that activate small molecule fluorophores have been selected in vitro. Among these, the ~30 nucleotide Mango-III is notable because it binds the thiazole orange derivative TO1-Biotin with high affinity and fluoresces brightly (quantum yield 0.55). Uniquely among related aptamers, Mango-III exhibits biphasic thermal melting, characteristic of molecules with tertiary structure. We report crystal structures of TO1-Biotin complexes of Mango-III, a structure-guided mutant Mango-III(A10U), and a functionally reselected mutant iMango-III. The structures reveal a globular architecture arising from an unprecedented pseudoknot-like connectivity between a G-quadruplex and an embedded non-canonical duplex. The fluorophore is restrained into a planar conformation by the G-quadruplex, a lone, long-range trans-Watson-Crick pair (whose A10U mutation increases quantum yield to 0.66), and a pyrimidine perpendicular to the nucleobase planes of those motifs. The improved iMango-III and Mango-III(A10U) fluoresce ~50% brighter than enhanced green fluorescent protein, making them suitable tags for live cell RNA visualization.
-
Structure and functional reselection of the Mango-III fluorogenic RNA aptamer.
Nature chemical biology, 2019Co-Authors: R.j. Trachoma, Alexis Autour, Amir Abdolahzadeh, Alessio Andreoni, Razvan Cojocaru, Ramil Garipov, Elena Dolgosheina, Sunny C. Y. Jeng, Jay R. Knutson, Michael RyckelynckAbstract:Several turn-on RNA aptamers that activate small-molecule fluorophores have been selected in vitro. Among these, the ~30 nucleotide Mango-III is notable because it binds the thiazole orange derivative TO1-Biotin with high affinity and fluoresces brightly (quantum yield 0.55). Uniquely among related aptamers, Mango-III exhibits biphasic thermal melting, characteristic of molecules with tertiary structure. We report crystal structures of TO1-Biotin complexes of Mango-III, a structure-guided mutant Mango-III(A10U), and a functionally reselected mutant iMango-III. The structures reveal a globular architecture arising from an unprecedented pseudoknot-like connectivity between a G-quadruplex and an embedded non-canonical duplex. The fluorophore is restrained into a planar conformation by the G-quadruplex, a lone, long-range trans Watson–Crick pair (whose A10U mutation increases quantum yield to 0.66), and a pyrimidine perpendicular to the nucleobase planes of those motifs. The improved iMango-III and Mango-III(A10U) fluoresce ~50% brighter than enhanced green fluorescent protein, making them suitable tags for live cell RNA visualization. Structural analysis of Mango-III in complex with fluorophores reveals a globular architecture and a planar conformation of fluorophores. Structure-guided mutation and functional reselection identified two mutants with improved fluorescence intensity.
-
Fluorogenic RNA Mango aptamers for imaging small non-coding RNAs in mammalian cells
Nature Communications, 2018Co-Authors: Alexis Autour, Sunny Jeng, Amir Abdolahzadeh, Michael Ryckelynck, Adam Cawte, Angela Galli, Shanker S. S. Panchapakesan, David Rueda, Peter J. UnrauAbstract:Despite having many key roles in cellular biology, directly imaging biologically important RNAs has been hindered by a lack of fluorescent tools equivalent to the fluorescent proteins available to study cellular proteins. Ideal RNA labelling systems must preserve biological function, have photophysical properties similar to existing fluorescent proteins, and be compatible with established live and fixed cell protein labelling strategies. Here, we report a microfluidics-based selection of three new high-affinity RNA Mango fluorogenic aptamers. Two of these are as bright or brighter than enhanced GFP when bound to TO1-Biotin. Furthermore, we show that the new Mangos can accurately image the subcellular localization of three small non-coding RNAs (5S, U6, and a box C/D scaRNA) in fixed and live mammalian cells. These new aptamers have many potential applications to study RNA function and dynamics both in vitro and in mammalian cells. Many aptamer-fluorophore complexes suffer from low quantum yield and low extinction coefficients limiting their usability. Here the authors isolate new Mango aptamers with improved fluorescent properties for both fixed and live-cell imaging by using competitive ligand binding microfluidic selection.
-
fluorogenic rna Mango aptamers for imaging small non coding rnas in mammalian cells
Nature Communications, 2018Co-Authors: Alexis Autour, Amir Abdolahzadeh, Michael Ryckelynck, Sunny C. Y. Jeng, Adam Cawte, Angela Galli, Shanker S. S. Panchapakesan, David Rueda, Peter J. UnrauAbstract:Despite having many key roles in cellular biology, directly imaging biologically important RNAs has been hindered by a lack of fluorescent tools equivalent to the fluorescent proteins available to study cellular proteins. Ideal RNA labelling systems must preserve biological function, have photophysical properties similar to existing fluorescent proteins, and be compatible with established live and fixed cell protein labelling strategies. Here, we report a microfluidics-based selection of three new high-affinity RNA Mango fluorogenic aptamers. Two of these are as bright or brighter than enhanced GFP when bound to TO1-Biotin. Furthermore, we show that the new Mangos can accurately image the subcellular localization of three small non-coding RNAs (5S, U6, and a box C/D scaRNA) in fixed and live mammalian cells. These new aptamers have many potential applications to study RNA function and dynamics both in vitro and in mammalian cells.
Carlos H. Crisosto - One of the best experts on this subject based on the ideXlab platform.
-
describing quality and sensory attributes of 3 Mango mangifera indica l cultivars at 3 ripeness stages based on firmness
Journal of Food Science, 2015Co-Authors: Rita De Cassia Mirela Resende Nassu, Eduardo Valério De Barros Vilas Boas, Luiz Carlos De Oliveira Lima, Sara Gonzalezmoscoso, Gayle M. Crisosto, Carlos H. CrisostoAbstract:To determine the ideal ripening stage for consumption of the Mango cultivars, "Ataulfo," "Haden," and "Tommy Atkins"; fruits at 3 flesh firmness levels (ripeness stages) were evaluated by a trained panel using descriptive analysis after instrumental measurements were made. After harvest, all fruits were ripened to allow softening and quality and sensory attribute changes. Ripening changes during softening of Ataulfo Mangos were expressed by a characteristic increase in the perception of "tropical fruit" and "peach" aromas, an increase in "juiciness," "sweetness," and "tropical fruit" flavor, while "fibrousness," "chewiness," and "sourness" decreased. Similar desirable sensory changes were also detected during softening of Haden Mangos; an increase in tropical fruit and peach aromas, sweetness and tropical fruit flavor, and a decrease in chewiness, sourness, and bitterness. Softening of Tommy Atkins Mangos was followed by reduced chewiness and sourness and increased peach aroma. Softening of all cultivars was followed by decreased sourness and titratable acidity (TA) and increased soluble solids concentration (SSC) and SSC:TA ratio. The results indicate that Mango ripening leads to increased expression of sensory attributes such as tropical fruit and peach aromas, tropical flavor, and sweetness that have been related to improved eating quality and these final changes in sensory quality attributes are specific for each cultivar. For example, Ataulfo and Haden Mangos had greater improvement in quality and sensory attributes related to fruit eating quality during ripening-softening than Tommy Atkins. In our consumer test, these quality-sensory attributes expressed during ripening that were perceived by the trained panel were also validated, supporting the need for a controlled ripening protocol in Mangos.
-
Describing Quality and Sensory Attributes of 3 Mango (Mangifera indica L.) Cultivars at 3 Ripeness Stages Based on Firmness.
Journal of Food Science, 2015Co-Authors: Rita De Cássia Mirela Resende Nassur, Sara González-moscoso, Eduardo Valério De Barros Vilas Boas, Luiz Carlos De Oliveira Lima, Gayle M. Crisosto, Carlos H. CrisostoAbstract:To determine the ideal ripening stage for consumption of the Mango cultivars, “Ataulfo,” “Haden,” and “Tommy Atkins”; fruits at 3 flesh firmness levels (ripeness stages) were evaluated by a trained panel using descriptive analysis after instrumental measurements were made. After harvest, all fruits were ripened to allow softening and quality and sensory attribute changes. Ripening changes during softening of Ataulfo Mangos were expressed by a characteristic increase in the perception of “tropical fruit” and “peach” aromas, an increase in “juiciness,” “sweetness,” and “tropical fruit” flavor, while “fibrousness,” “chewiness,” and “sourness” decreased. Similar desirable sensory changes were also detected during softening of Haden Mangos; an increase in tropical fruit and peach aromas, sweetness and tropical fruit flavor, and a decrease in chewiness, sourness, and bitterness. Softening of Tommy Atkins Mangos was followed by reduced chewiness and sourness and increased peach aroma. Softening of all cultivars was followed by decreased sourness and titratable acidity (TA) and increased soluble solids concentration (SSC) and SSC:TA ratio. The results indicate that Mango ripening leads to increased expression of sensory attributes such as tropical fruit and peach aromas, tropical flavor, and sweetness that have been related to improved eating quality and these final changes in sensory quality attributes are specific for each cultivar. For example, Ataulfo and Haden Mangos had greater improvement in quality and sensory attributes related to fruit eating quality during ripening-softening than Tommy Atkins. In our consumer test, these quality-sensory attributes expressed during ripening that were perceived by the trained panel were also validated, supporting the need for a controlled ripening protocol in Mangos. Practical Application This research provides information on the benefits of ripening on Mango sensory attributes identified as important to improve eating quality of the fruit. With this information, postharvest handling and cultivar characteristics of Mangos can be improved to increase consumer satisfaction.
