Q3138236 (Q3138236): Difference between revisions
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(Created claim: summary (P836): ARGONAUTA PROTEINS PRODUCE RNA-RNA INTERFERENCE RESPONSIBLE FOR GENIC SILENCING IN EUKARYOTES, ALTHOUGH MANY ARCHAEA AND SOME BACTERIA ALSO ENCODE PROTEINS FROM THE ARGONAUT FAMILY. THE THAGO PROTEIN FROM THERMUS THERMOPHILUS (HUMAN AGO2 PROTEIN HORMONE) PRESENTS THE UNUSUAL ABILITY TO CUT DNA COMPLEMENTARY TO SSDNA GUIDES, WHICH IS A NOVEL DNA-DNA INTERFERENCE SYSTEM. PREVIOUS DATA FROM THE REQUESTING LABORATORY SHOW THAT THAGO ACTS AS A BARRIE...) |
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ARGONAUTA PROTEINS PRODUCE RNA-RNA INTERFERENCE RESPONSIBLE FOR GENIC SILENCING IN EUKARYOTES, ALTHOUGH MANY ARCHAEA AND SOME BACTERIA ALSO ENCODE PROTEINS FROM THE ARGONAUT FAMILY. THE THAGO PROTEIN FROM THERMUS THERMOPHILUS (HUMAN AGO2 PROTEIN HORMONE) PRESENTS THE UNUSUAL ABILITY TO CUT DNA COMPLEMENTARY TO SSDNA GUIDES, WHICH IS A NOVEL DNA-DNA INTERFERENCE SYSTEM. PREVIOUS DATA FROM THE REQUESTING LABORATORY SHOW THAT THAGO ACTS AS A BARRIER OF INNATE IMMUNITY AGAINST EXOGENOUS DNA, EFFECTIVELY DESTROYING MOST (90 %) OF THE DNA MOLECULES THAT ENTER THE CELL BY TRANSFORMATION, BUT WITHOUT ACTING ON THE SAME DNA WHEN IT IS TRANSFERRED FROM ANOTHER BACTERIUM OF THE SAME GENUS, SUGGESTING THAT THAGO’S ACTIVITY IS SUBJECT TO REGULATION IN THE CELL. ON THE OTHER HAND, THE CUTTING CAPACITY LED BY SSDNA OF THAGO CONSTITUTES A PROPERTY OF EXTRAORDINARY APPLICATION POTENTIAL IN GENOME EDITION, SIMILAR IN ITS OPERATION BUT WITH POTENTIAL ADVANTAGES OVER THE CRISPR/CAS9 SYSTEM. IN FACT, IN A PROJECT EXPLORED COMPLETED IN 2015 WE OBSERVED THAT THE TRANSIENT EXPRESSION OF THAGO IN TRANSFECTED RATON CELLS WITH SPECIFIC OLIGONUCLEOTIDES PRODUCED THE SHUTDOWN IN THE EXPRESSION OF GFP FROM A PLASMAID. In THE PROJECT THAT IS REQUESTed we intend to PROFUND in the MECHANISMS that allow you to make it available to you to enter DNA ADQUIRIED by TRANSFORMATION or TRANSFERIING OF Other CELULS WITH THE FINANCIAL AND OPTIMITION OF YOUR FUNCTIONING OF HIS APPLICATION AS TOOL IN THE EDITION OF GENOMAS._x000D_ as a preliminary step to any process of openness of the PROTEIN, THE FIRST OBJECTIVE PROPOSAL PROPOSAL IENTIFICATION OF COMPONENTS AND ACTIVITIES/deactivation MECANISMs included in the DNA-DNA Interference Answer Measured by THAGO, it will be addressed by means of a combination of biochemical, protheomic and GENETIC technologies for identification of covalent modifications and interactions with other necessities for interference. IN THE SECOND OBJECTIVE, WILL THE USE OF PROTEINS PAID AS TOOLS FOR DNA MODIFICATION BE ADDRESSED THE MODIFICATION OF THAGO FOCUSED ON THE DATA OBTAINED IN THE FIRST OBJECTIVE, AS WELL AS RANDOMLY, THROUGH IN VITRO DIRECTED EVOLUTION. THE MOST EFFICIENT VARIANTS IN CUTTING TESTS LED BY GUIA WILL BE USED IN VITRO IN DNA MODIFICATION PROCESSES AS A PROGRAMMABLE RESTRICTION ENZYME, AND APPLIED DIRECTLY IN VIVO FOR THE EDITING OF GENOMES OF PROKARYOTES AND EUKARYOTES. IN THE CASE OF GENOMES OF EUKARYOTES, THE ADDITION OF NUCLEAR LOCATION SIGNALS TO THESE IMPROVED VARIANTS WOULD ALLOW DIRECT TRANSFECCION OF THE PROTEIN ALREADY LINKED TO SSDNA GUIDES, AVOIDING THE RISK OF ACCIDENTAL INCORPORATION OF EXOGENOUS DNA INTO THE GENOME ASSOCIATED WITH THE CURRENT CRISPR/CAS9 TECHNIQUES. (English) | |||||||||||||||
Property / summary: ARGONAUTA PROTEINS PRODUCE RNA-RNA INTERFERENCE RESPONSIBLE FOR GENIC SILENCING IN EUKARYOTES, ALTHOUGH MANY ARCHAEA AND SOME BACTERIA ALSO ENCODE PROTEINS FROM THE ARGONAUT FAMILY. THE THAGO PROTEIN FROM THERMUS THERMOPHILUS (HUMAN AGO2 PROTEIN HORMONE) PRESENTS THE UNUSUAL ABILITY TO CUT DNA COMPLEMENTARY TO SSDNA GUIDES, WHICH IS A NOVEL DNA-DNA INTERFERENCE SYSTEM. PREVIOUS DATA FROM THE REQUESTING LABORATORY SHOW THAT THAGO ACTS AS A BARRIER OF INNATE IMMUNITY AGAINST EXOGENOUS DNA, EFFECTIVELY DESTROYING MOST (90 %) OF THE DNA MOLECULES THAT ENTER THE CELL BY TRANSFORMATION, BUT WITHOUT ACTING ON THE SAME DNA WHEN IT IS TRANSFERRED FROM ANOTHER BACTERIUM OF THE SAME GENUS, SUGGESTING THAT THAGO’S ACTIVITY IS SUBJECT TO REGULATION IN THE CELL. ON THE OTHER HAND, THE CUTTING CAPACITY LED BY SSDNA OF THAGO CONSTITUTES A PROPERTY OF EXTRAORDINARY APPLICATION POTENTIAL IN GENOME EDITION, SIMILAR IN ITS OPERATION BUT WITH POTENTIAL ADVANTAGES OVER THE CRISPR/CAS9 SYSTEM. IN FACT, IN A PROJECT EXPLORED COMPLETED IN 2015 WE OBSERVED THAT THE TRANSIENT EXPRESSION OF THAGO IN TRANSFECTED RATON CELLS WITH SPECIFIC OLIGONUCLEOTIDES PRODUCED THE SHUTDOWN IN THE EXPRESSION OF GFP FROM A PLASMAID. In THE PROJECT THAT IS REQUESTed we intend to PROFUND in the MECHANISMS that allow you to make it available to you to enter DNA ADQUIRIED by TRANSFORMATION or TRANSFERIING OF Other CELULS WITH THE FINANCIAL AND OPTIMITION OF YOUR FUNCTIONING OF HIS APPLICATION AS TOOL IN THE EDITION OF GENOMAS._x000D_ as a preliminary step to any process of openness of the PROTEIN, THE FIRST OBJECTIVE PROPOSAL PROPOSAL IENTIFICATION OF COMPONENTS AND ACTIVITIES/deactivation MECANISMs included in the DNA-DNA Interference Answer Measured by THAGO, it will be addressed by means of a combination of biochemical, protheomic and GENETIC technologies for identification of covalent modifications and interactions with other necessities for interference. IN THE SECOND OBJECTIVE, WILL THE USE OF PROTEINS PAID AS TOOLS FOR DNA MODIFICATION BE ADDRESSED THE MODIFICATION OF THAGO FOCUSED ON THE DATA OBTAINED IN THE FIRST OBJECTIVE, AS WELL AS RANDOMLY, THROUGH IN VITRO DIRECTED EVOLUTION. THE MOST EFFICIENT VARIANTS IN CUTTING TESTS LED BY GUIA WILL BE USED IN VITRO IN DNA MODIFICATION PROCESSES AS A PROGRAMMABLE RESTRICTION ENZYME, AND APPLIED DIRECTLY IN VIVO FOR THE EDITING OF GENOMES OF PROKARYOTES AND EUKARYOTES. IN THE CASE OF GENOMES OF EUKARYOTES, THE ADDITION OF NUCLEAR LOCATION SIGNALS TO THESE IMPROVED VARIANTS WOULD ALLOW DIRECT TRANSFECCION OF THE PROTEIN ALREADY LINKED TO SSDNA GUIDES, AVOIDING THE RISK OF ACCIDENTAL INCORPORATION OF EXOGENOUS DNA INTO THE GENOME ASSOCIATED WITH THE CURRENT CRISPR/CAS9 TECHNIQUES. (English) / rank | |||||||||||||||
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Property / summary: ARGONAUTA PROTEINS PRODUCE RNA-RNA INTERFERENCE RESPONSIBLE FOR GENIC SILENCING IN EUKARYOTES, ALTHOUGH MANY ARCHAEA AND SOME BACTERIA ALSO ENCODE PROTEINS FROM THE ARGONAUT FAMILY. THE THAGO PROTEIN FROM THERMUS THERMOPHILUS (HUMAN AGO2 PROTEIN HORMONE) PRESENTS THE UNUSUAL ABILITY TO CUT DNA COMPLEMENTARY TO SSDNA GUIDES, WHICH IS A NOVEL DNA-DNA INTERFERENCE SYSTEM. PREVIOUS DATA FROM THE REQUESTING LABORATORY SHOW THAT THAGO ACTS AS A BARRIER OF INNATE IMMUNITY AGAINST EXOGENOUS DNA, EFFECTIVELY DESTROYING MOST (90 %) OF THE DNA MOLECULES THAT ENTER THE CELL BY TRANSFORMATION, BUT WITHOUT ACTING ON THE SAME DNA WHEN IT IS TRANSFERRED FROM ANOTHER BACTERIUM OF THE SAME GENUS, SUGGESTING THAT THAGO’S ACTIVITY IS SUBJECT TO REGULATION IN THE CELL. ON THE OTHER HAND, THE CUTTING CAPACITY LED BY SSDNA OF THAGO CONSTITUTES A PROPERTY OF EXTRAORDINARY APPLICATION POTENTIAL IN GENOME EDITION, SIMILAR IN ITS OPERATION BUT WITH POTENTIAL ADVANTAGES OVER THE CRISPR/CAS9 SYSTEM. IN FACT, IN A PROJECT EXPLORED COMPLETED IN 2015 WE OBSERVED THAT THE TRANSIENT EXPRESSION OF THAGO IN TRANSFECTED RATON CELLS WITH SPECIFIC OLIGONUCLEOTIDES PRODUCED THE SHUTDOWN IN THE EXPRESSION OF GFP FROM A PLASMAID. In THE PROJECT THAT IS REQUESTed we intend to PROFUND in the MECHANISMS that allow you to make it available to you to enter DNA ADQUIRIED by TRANSFORMATION or TRANSFERIING OF Other CELULS WITH THE FINANCIAL AND OPTIMITION OF YOUR FUNCTIONING OF HIS APPLICATION AS TOOL IN THE EDITION OF GENOMAS._x000D_ as a preliminary step to any process of openness of the PROTEIN, THE FIRST OBJECTIVE PROPOSAL PROPOSAL IENTIFICATION OF COMPONENTS AND ACTIVITIES/deactivation MECANISMs included in the DNA-DNA Interference Answer Measured by THAGO, it will be addressed by means of a combination of biochemical, protheomic and GENETIC technologies for identification of covalent modifications and interactions with other necessities for interference. IN THE SECOND OBJECTIVE, WILL THE USE OF PROTEINS PAID AS TOOLS FOR DNA MODIFICATION BE ADDRESSED THE MODIFICATION OF THAGO FOCUSED ON THE DATA OBTAINED IN THE FIRST OBJECTIVE, AS WELL AS RANDOMLY, THROUGH IN VITRO DIRECTED EVOLUTION. THE MOST EFFICIENT VARIANTS IN CUTTING TESTS LED BY GUIA WILL BE USED IN VITRO IN DNA MODIFICATION PROCESSES AS A PROGRAMMABLE RESTRICTION ENZYME, AND APPLIED DIRECTLY IN VIVO FOR THE EDITING OF GENOMES OF PROKARYOTES AND EUKARYOTES. IN THE CASE OF GENOMES OF EUKARYOTES, THE ADDITION OF NUCLEAR LOCATION SIGNALS TO THESE IMPROVED VARIANTS WOULD ALLOW DIRECT TRANSFECCION OF THE PROTEIN ALREADY LINKED TO SSDNA GUIDES, AVOIDING THE RISK OF ACCIDENTAL INCORPORATION OF EXOGENOUS DNA INTO THE GENOME ASSOCIATED WITH THE CURRENT CRISPR/CAS9 TECHNIQUES. (English) / qualifier | |||||||||||||||
point in time: 12 October 2021
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Revision as of 12:33, 12 October 2021
Project Q3138236 in Spain
Language | Label | Description | Also known as |
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English | No label defined |
Project Q3138236 in Spain |
Statements
108,900.0 Euro
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217,800.0 Euro
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50.0 percent
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30 December 2016
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31 December 2020
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UNIVERSIDAD AUTONOMA DE MADRID
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28079
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LAS PROTEINAS ARGONAUTA PRODUCEN LA INTERFERENCIA RNA-RNA RESPONSABLE DEL SILENCIAMIENTO GENICO EN EUCARIOTAS, AUNQUE MUCHAS ARQUEAS Y ALGUNAS BACTERIAS TAMBIEN CODIFICAN PROTEINAS DE LA FAMILIA ARGONAUTA. LA PROTEINA THAGO DE THERMUS THERMOPHILUS ( HOMOLOGA DE LA PROTEINA AGO2 HUMANA) PRESENTA LA INUSUAL CAPACIDAD DE CORTAR DNA COMPLEMENTARIO A GUIAS DE SSDNA, LO QUE CONSTITUYE UN NOVEDOSO SISTEMA DE INTERFERENCIA DNA-DNA. DATOS PREVIOS DEL LABORATORIO SOLICITANTE MUESTRAN QUE THAGO ACTUA COMO BARRERA DE INMUNIDAD INNATA FRENTE A DNA EXOGENO, DESTRUYENDO DE FORMA EFICAZ LA MAYOR PARTE (90%) DE LAS MOLECULAS DE DNA QUE ENTRAN EN LA CELULA POR TRANSFORMACION, PERO SIN ACTUAR SOBRE EL MISMO DNA CUANDO ESTE ES TRANSFERIDO DESDE OTRA BACTERIA DEL MISMO GENERO, LO QUE SUGIERE QUE LA ACTIVIDAD DE THAGO ESTA SOMETIDA A REGULACION EN LA CELULA. POR OTRA PARTE, LA CAPACIDAD DE CORTE DIRIGIDO POR SSDNA DE THAGO CONSTITUYE UNA PROPIEDAD DE EXTRAORDINARIO POTENCIAL DE APLICACION EN EDICION DE GENOMAS, SIMILAR EN SU FUNCIONAMIENTO PERO CON POTENCIALES VENTAJAS SOBRE EL SISTEMA CRISPR/CAS9. DE HECHO, EN UN PROYECTO EXPLORA FINALIZADO EN 2015 OBSERVAMOS QUE LA EXPRESION TRANSITORIA DE THAGO EN CELULAS DE RATON TRANSFECTADAS CON OLIGONUCLEOTIDOS GUIA ESPECIFICOS PRODUCIA EL APAGADO EN LA EXPRESION DE GFP DESDE UN PLASMIDO. EN EL PROYECTO QUE SE SOLICITA PRETENDEMOS PROFUNDIZAR EN LOS MECANISMOS QUE LE PERMITEN A THAGO DISTINGUIR ENTRE DNA ADQUIRIDO POR TRANSFORMACION O TRANSFERIDO DE OTRAS CELULAS CON EL FIN DE ENTENDER Y OPTIMIZAR SU FUNCIONAMIENTO DE CARA A SU APLICACION COMO HERRAMIENTA EN LA EDICION DE GENOMAS._x000D_ COMO PASO PREVIO A CUALQUIER PROCESO DE OPTIMIZACION DE LA PROTEINA, EL PRIMER OBJETIVO PROPUESTO TITULADO ¿IDENTIFICACION DE LOS COMPONENTES Y LOS MECANISMOS DE ACTIVACION/DESACTIVACION IMPLICADOS EN LA RESPUESTA DE INTERFERENCIA DNA-DNA MEDIADA POR THAGO¿, SERA ABORDADO MEDIANTE UNA COMBINACION DE TECNICAS BIOQUIMICAS, PROTEOMICAS Y GENETICAS PARA IDENTIFICAR POSIBLES MODIFICACIONES COVALENTES E INTERACCIONES CON OTRAS PROTEINAS NECESARIAS PARA LA INTERFERENCIA. EN EL SEGUNDO OBJETIVO ¿UTILIZACION DE PROTEINAS PAGO COMO HERRAMIENTAS PARA LA MODIFICACION DE DNA¿, SE ABORDARA LA MODIFICACION DE THAGO TANTO FOCALIZADA EN BASE A LOS DATOS OBTENIDOS EN EL PRIMER OBJETIVO, COMO AL AZAR, MEDIANTE EVOLUCION DIRIGIDA IN VITRO. LAS VARIANTES MAS EFICIENTES EN ENSAYOS DE CORTE DIRIGIDOS POR GUIA SERAN UTILIZADAS IN VITRO EN PROCESOS DE MODIFICACION DE DNA COMO ENZIMA DE RESTRICCION PROGRAMABLE, Y APLICADAS DIRECTAMENTE IN VIVO PARA LA EDICION DE GENOMAS DE PROCARIOTAS Y EUCARIOTAS. EN EL CASO DE LOS GENOMAS DE EUCARIOTAS, LA ADICION DE DE SEÑALES DE LOCALIZACION NUCLEAR A ESTAS VARIANTES MEJORADAS PERMITIRIA LA TRANSFECCION DIRECTA DE LA PROTEINA YA UNIDA A GUIAS DE SSDNA, EVITANDO EL RIESGO DE INCORPORACION ACCIDENTAL DE DNA EXOGENO EN EL GENOMA ASOCIADO A LAS TECNICAS CRISPR/CAS9 ACTUALES. (Spanish)
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ARGONAUTA PROTEINS PRODUCE RNA-RNA INTERFERENCE RESPONSIBLE FOR GENIC SILENCING IN EUKARYOTES, ALTHOUGH MANY ARCHAEA AND SOME BACTERIA ALSO ENCODE PROTEINS FROM THE ARGONAUT FAMILY. THE THAGO PROTEIN FROM THERMUS THERMOPHILUS (HUMAN AGO2 PROTEIN HORMONE) PRESENTS THE UNUSUAL ABILITY TO CUT DNA COMPLEMENTARY TO SSDNA GUIDES, WHICH IS A NOVEL DNA-DNA INTERFERENCE SYSTEM. PREVIOUS DATA FROM THE REQUESTING LABORATORY SHOW THAT THAGO ACTS AS A BARRIER OF INNATE IMMUNITY AGAINST EXOGENOUS DNA, EFFECTIVELY DESTROYING MOST (90 %) OF THE DNA MOLECULES THAT ENTER THE CELL BY TRANSFORMATION, BUT WITHOUT ACTING ON THE SAME DNA WHEN IT IS TRANSFERRED FROM ANOTHER BACTERIUM OF THE SAME GENUS, SUGGESTING THAT THAGO’S ACTIVITY IS SUBJECT TO REGULATION IN THE CELL. ON THE OTHER HAND, THE CUTTING CAPACITY LED BY SSDNA OF THAGO CONSTITUTES A PROPERTY OF EXTRAORDINARY APPLICATION POTENTIAL IN GENOME EDITION, SIMILAR IN ITS OPERATION BUT WITH POTENTIAL ADVANTAGES OVER THE CRISPR/CAS9 SYSTEM. IN FACT, IN A PROJECT EXPLORED COMPLETED IN 2015 WE OBSERVED THAT THE TRANSIENT EXPRESSION OF THAGO IN TRANSFECTED RATON CELLS WITH SPECIFIC OLIGONUCLEOTIDES PRODUCED THE SHUTDOWN IN THE EXPRESSION OF GFP FROM A PLASMAID. In THE PROJECT THAT IS REQUESTed we intend to PROFUND in the MECHANISMS that allow you to make it available to you to enter DNA ADQUIRIED by TRANSFORMATION or TRANSFERIING OF Other CELULS WITH THE FINANCIAL AND OPTIMITION OF YOUR FUNCTIONING OF HIS APPLICATION AS TOOL IN THE EDITION OF GENOMAS._x000D_ as a preliminary step to any process of openness of the PROTEIN, THE FIRST OBJECTIVE PROPOSAL PROPOSAL IENTIFICATION OF COMPONENTS AND ACTIVITIES/deactivation MECANISMs included in the DNA-DNA Interference Answer Measured by THAGO, it will be addressed by means of a combination of biochemical, protheomic and GENETIC technologies for identification of covalent modifications and interactions with other necessities for interference. IN THE SECOND OBJECTIVE, WILL THE USE OF PROTEINS PAID AS TOOLS FOR DNA MODIFICATION BE ADDRESSED THE MODIFICATION OF THAGO FOCUSED ON THE DATA OBTAINED IN THE FIRST OBJECTIVE, AS WELL AS RANDOMLY, THROUGH IN VITRO DIRECTED EVOLUTION. THE MOST EFFICIENT VARIANTS IN CUTTING TESTS LED BY GUIA WILL BE USED IN VITRO IN DNA MODIFICATION PROCESSES AS A PROGRAMMABLE RESTRICTION ENZYME, AND APPLIED DIRECTLY IN VIVO FOR THE EDITING OF GENOMES OF PROKARYOTES AND EUKARYOTES. IN THE CASE OF GENOMES OF EUKARYOTES, THE ADDITION OF NUCLEAR LOCATION SIGNALS TO THESE IMPROVED VARIANTS WOULD ALLOW DIRECT TRANSFECCION OF THE PROTEIN ALREADY LINKED TO SSDNA GUIDES, AVOIDING THE RISK OF ACCIDENTAL INCORPORATION OF EXOGENOUS DNA INTO THE GENOME ASSOCIATED WITH THE CURRENT CRISPR/CAS9 TECHNIQUES. (English)
12 October 2021
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Madrid
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Identifiers
BIO2016-77031-R
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