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| Acceso al texto completo restringido a Biblioteca INIA La Estanzuela. Por información adicional contacte bib_le@inia.org.uy. |
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Biblioteca (s) : |
INIA La Estanzuela. |
Fecha : |
12/07/2022 |
Actualizado : |
12/07/2022 |
Tipo de producción científica : |
Capítulo en Libro Técnico-Científico |
Autor : |
KALDS, P.; CRISPO, M.; TESSON, L.; ANEGÓN, I.; CHEN KEY, Y.; WANG, X.; MENCHACA, A. |
Afiliación : |
PETER KALDS, Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, China.; MARTINA CRISPO, Unidad de Biotecnología en Animales de Laboratorio (UBAL), Institut Pasteur de Montevideo, Montevideo, Uruguay.; LAURENT TESSON, INSERM, Centre de Recherche en Transplantation et Immunologie, UMR 1064, Transgenesis Rat ImmunoPhenomic Facility (TRIP), Nantes, France.; IGNACIO ANEGÓN, INSERM, Centre de Recherche en Transplantation et Immunologie, UMR 1064, Nantes, France.; YULIN CHEN KEY, Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, China.; XIAOLONG WANG, Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, China.; JOSE ALEJO MENCHACA BARBEITO, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay./Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay. |
Título : |
Generation of Double-Muscled Sheep and Goats by CRISPR /Cas9-Mediated Knockout of the Myostatin Gene. |
Complemento del título : |
Chapter 16. |
Fecha de publicación : |
2022 |
Fuente / Imprenta : |
Methods in Molecular Biology, 2022, Volume 2495, Pages 295-323. Doi: https://doi.org/10.1007/978-1-0716-2301-5_16 |
ISBN : |
Online: 978-1-0716-2301-5 |
DOI : |
10.1007/978-1-0716-2301-5_16 |
Idioma : |
Inglés |
Contenido : |
Abstract:
The myostatin (MSTN) gene has shown to play a critical role in the regulation of skeletal muscle mass, and the translational inhibition of this gene has shown increased muscle mass, generating what is known as ?double-muscling phenotype.? Disruption of the MSTN gene expression using the CRISPR/Cas9 genome-editing system has shown improved muscle development and growth rates in livestock species, including sheep and goats. Here, we describe procedures for the generation of MSTN knockout sheep and goats using the microinjection approach of the CRISPR/Cas9 system, including the selection of targeting sgRNAs, the construction of CRISPR/Cas9 targeting vector, the in vitro examination of system efficiency, the in vivo targeting to generate MSTN knockout founders, the genomic and phenotypic characterization of the generated offspring, and the assessment of off-target effects in gene-edited founders through targeted validation of predicted off-target sites, as well as genome-wide off-target analysis by whole-genome sequencing. Editing the MSTN gene using the CRISPR/Cas9 system might be a rapid and promising alternative to promote meat production in livestock. |
Palabras claves : |
CRISPR/Cas9; Genome editing; Goats; Knockout; Meat production; Microinjection; MSTN; Sheep; Small ruminants. |
Thesagro : |
CABRAS; OVEJA; PRODUCCION DE CARNE. |
Asunto categoría : |
-- |
Marc : |
LEADER 02176naa a2200349 a 4500 001 1063414 005 2022-07-12 008 2022 bl uuuu u00u1 u #d 024 7 $a10.1007/978-1-0716-2301-5_16$2DOI 100 1 $aKALDS, P. 245 $aGeneration of Double-Muscled Sheep and Goats by CRISPR /Cas9-Mediated Knockout of the Myostatin Gene.$h[electronic resource] 260 $c2022 520 $aAbstract: The myostatin (MSTN) gene has shown to play a critical role in the regulation of skeletal muscle mass, and the translational inhibition of this gene has shown increased muscle mass, generating what is known as ?double-muscling phenotype.? Disruption of the MSTN gene expression using the CRISPR/Cas9 genome-editing system has shown improved muscle development and growth rates in livestock species, including sheep and goats. Here, we describe procedures for the generation of MSTN knockout sheep and goats using the microinjection approach of the CRISPR/Cas9 system, including the selection of targeting sgRNAs, the construction of CRISPR/Cas9 targeting vector, the in vitro examination of system efficiency, the in vivo targeting to generate MSTN knockout founders, the genomic and phenotypic characterization of the generated offspring, and the assessment of off-target effects in gene-edited founders through targeted validation of predicted off-target sites, as well as genome-wide off-target analysis by whole-genome sequencing. Editing the MSTN gene using the CRISPR/Cas9 system might be a rapid and promising alternative to promote meat production in livestock. 650 $aCABRAS 650 $aOVEJA 650 $aPRODUCCION DE CARNE 653 $aCRISPR/Cas9 653 $aGenome editing 653 $aGoats 653 $aKnockout 653 $aMeat production 653 $aMicroinjection 653 $aMSTN 653 $aSheep 653 $aSmall ruminants 700 1 $aCRISPO, M. 700 1 $aTESSON, L. 700 1 $aANEGÓN, I. 700 1 $aCHEN KEY, Y. 700 1 $aWANG, X. 700 1 $aMENCHACA, A. 773 $tMethods in Molecular Biology, 2022, Volume 2495, Pages 295-323. Doi: https://doi.org/10.1007/978-1-0716-2301-5_16
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| Acceso al texto completo restringido a Biblioteca INIA Las Brujas. Por información adicional contacte bibliolb@inia.org.uy. |
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Biblioteca (s) : |
INIA Las Brujas. |
Fecha actual : |
17/05/2022 |
Actualizado : |
02/12/2022 |
Tipo de producción científica : |
Artículos en Revistas Indexadas Internacionales |
Circulación / Nivel : |
Internacional - -- |
Autor : |
PASSOS, J. R. S.; GUERREIRO, D. D.; OTÁVIO, K. S.; SANTOS-NETO, P. C. DOS; SOUZA-NEVES, M.; CUADRO, F.; NUÑEZ-OLIVERA, R.; CRISPO, M.; BEZERRA, M. J. B.; SILVA, R. F.; LIMA, L. F.; FIGUEIREDO, J. R.; BUSTAMANTE-FILHO, I. C.; MENCHACA, A.; MOURA, A. A. |
Afiliación : |
JOSÉ RENATO S. PASSOS, Laboratório de Fisiologia e Ciências Ômicas, Departamento de Zootecnia, Universidade Federal do Ceará, Fortaleza, Brazil; DENISE D. GUERREIRO, Laboratório de Fisiologia e Ciências Ômicas, Departamento de Zootecnia, Universidade Federal do Ceará, Fortaleza, Brazil; KAMILA S. OTÁVIO, Laboratório de Fisiologia e Ciências Ômicas, Departamento de Zootecnia, Universidade Federal do Ceará, Fortaleza, Brazil; P. C. DOS SANTOS-NETO, Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; MARCELA SOUZA-NEVES, Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; FEDERICO CUADRO, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; RICHARD NUÑEZ-OLIVERA, Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; MARTINA CRISPO, Unidad de Biotecnología en Animales de Laboratorio, Institut Pasteur de Montevideo, Montevideo, Uruguay; MARIA JÚLIA B. BEZERRA, Laboratório de Fisiologia e Ciências Ômicas, Departamento de Zootecnia, Universidade Federal do Ceará, Fortaleza, Brazil; RENATO F. SILVA, Laboratório de Manipulação de Oócitos e Folículos Ovarianos Pré-antrais - LAMOFOPA - Faculdade de Veterinária, Universidade Estadual do Ceará, Fortaleza, Brazil; LARITZA F. LIMA, Laboratório de Manipulação de Oócitos e Folículos Ovarianos Pré-antrais - LAMOFOPA - Faculdade de Veterinária, Universidade Estadual do Ceará, Fortaleza, Brazil; JOSÉ RICARDO FIGUEIREDO, Laboratório de Manipulação de Oócitos e Folículos Ovarianos Pré-antrais - LAMOFOPA - Faculdade de Veterinária, Universidade Estadual do Ceará, Fortaleza, Brazil; IVAN C. BUSTAMANTE-FILHO, aboratório de Biotecnologia da Reprodução Animal, Programa de Pós-graduação em Biotecnologia, Universidade do Vale do Taquari, Lajeado, Brazil; JOSE ALEJO MENCHACA BARBEITO, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; ARLINDO A. MOURA, Laboratório de Fisiologia e Ciências Ômicas, Departamento de Zootecnia, Universidade Federal do Ceará, Fortaleza, Brazil. |
Título : |
Global proteomic analysis of preimplantational ovine embryos produced in vitro. |
Fecha de publicación : |
2022 |
Fuente / Imprenta : |
Reproduction in Domestic Animals, 2022, Volume 57, Issue 7; pages 784-797. doi: https://doi.org/10.1111/rda.14122 |
ISSN : |
0936-6768 |
DOI : |
10.1111/rda.14122 |
Idioma : |
Inglés |
Notas : |
Article history: Received 15 February 2022; Accepted 1 April 2022. -- Funding text - The experiments presently described were conducted at the facilities of the (Fundacion IRAUy, Montevideo, Uruguay) and at the (UBAL) of the , Uruguay. Specially, the authors thank Dr. Rosario Durán and Dr. Alejandro Leyva for kindly assisting us in the proteomic experiment. Finnacial support was provided by Fundacion IRAUy; PRONEX 02/2015 (Programa de Apoio a Núcleos de Excelência Pronex/Funcap/CNPq); the Brazilian Research Council?CNPq (grants # 313160/2017‐1 and 438773/2018‐7); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brazil. Instituto de Reproducción Animal Uruguay Unidad de Biotecnología en Animales de Laboratorio Institut Pasteur de Montevideo. -- Corresponding author: A. Moura, A.; Laboratório de Fisiologia e Ciências Ômicas, Departamento de Zootecnia, Universidade Federal do Ceará, Fortaleza, Brazil; email:arlindo.moura@gmail.com -- Menchaca, A.; Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; mail:menchaca.alejo@gmail.com |
Contenido : |
ABSTRACT. -The present study was conducted to characterize the major proteome of preimplantation (D6) ovine embryos produced in vitro. COCs were aspirated from antral follicles (2–6 mm), matured and fertilized in vitro and cultured until day six. Proteins were ex- tracted separately from three pools of 45 embryos and separately run in SDS-PAGE. Proteins from each pool were individually subjected to in-gel digestion followed by LC-MS/MS. Three ‘raw files’ and protein lists were produced by Pattern Lab software, but only proteins present in all three lists were used for the bioinformatics analyses. There were 2,262 proteins identified in the 6-day-old ovine
embryos, including al- bumin, zona pellucida glycoprotein 2, 3 and 4, peptidyl arginine deiminase 6, actin cytoplasmic 1, gamma-actin 1, pyruvate kinase, heat shock protein 90 and protein disulfide isomerase, among others. Major biological processes linked to the sheep embryo proteome were translation, protein transport and protein stabilization, and molecular functions, defined as ATP binding, oxygen carrier activity and oxygen bind- ing. There were 42 enriched functional clusters
according to the 2,147 genes (UniProt database). Ten selected clusters with potential association with embryo development included translation, structural constituent of ribosomes, ribosomes, nucleosomes, structural constituent of the cytoskeleton, microtubule-based process, translation initiation factor activity, regulation of translational initiation, cell body and nucleotide biosynthetic process. The most representative KEEG pathways were ribosome, oxida- tive phosphorylation, glutathione metabolism, gap junction, mineral absorption, DNA replication and cGMP-PKG signalling pathway. Analyses of functional clusters clearly showed differences associated
with the proteome of preimplantation (D6) sheep em- bryos generated after in vitro fertilization in comparison with in vivo counterparts (Sanchez et al., 2021; https://doi.org/10.1111/rda.13897), confirming that the quality of in vitro derived blastocysts are unlike those produced in vivo. The present study portrays the first comprehensive overview of the proteome of preimplantational ovine embryos grown in vitro.
© 2022 Wiley-VCH GmbH. MenosABSTRACT. -The present study was conducted to characterize the major proteome of preimplantation (D6) ovine embryos produced in vitro. COCs were aspirated from antral follicles (2–6 mm), matured and fertilized in vitro and cultured until day six. Proteins were ex- tracted separately from three pools of 45 embryos and separately run in SDS-PAGE. Proteins from each pool were individually subjected to in-gel digestion followed by LC-MS/MS. Three ‘raw files’ and protein lists were produced by Pattern Lab software, but only proteins present in all three lists were used for the bioinformatics analyses. There were 2,262 proteins identified in the 6-day-old ovine
embryos, including al- bumin, zona pellucida glycoprotein 2, 3 and 4, peptidyl arginine deiminase 6, actin cytoplasmic 1, gamma-actin 1, pyruvate kinase, heat shock protein 90 and protein disulfide isomerase, among others. Major biological processes linked to the sheep embryo proteome were translation, protein transport and protein stabilization, and molecular functions, defined as ATP binding, oxygen carrier activity and oxygen bind- ing. There were 42 enriched functional clusters
according to the 2,147 genes (UniProt database). Ten selected clusters with potential association with embryo development included translation, structural constituent of ribosomes, ribosomes, nucleosomes, structural constituent of the cytoskeleton, microtubule-based process, translation initiation factor activity, regulation of translational i... Presentar Todo |
Palabras claves : |
Embryo development; In vitro fertilization; Mass spectrometry; Oocyte; Ovine; PLATAFORMA SALUD ANIMAL; Proteins. |
Asunto categoría : |
L10 Genética y mejoramiento animal |
Marc : |
LEADER 04571naa a2200409 a 4500 001 1063149 005 2022-12-02 008 2022 bl uuuu u00u1 u #d 022 $a0936-6768 024 7 $a10.1111/rda.14122$2DOI 100 1 $aPASSOS, J. R. S. 245 $aGlobal proteomic analysis of preimplantational ovine embryos produced in vitro.$h[electronic resource] 260 $c2022 500 $aArticle history: Received 15 February 2022; Accepted 1 April 2022. -- Funding text - The experiments presently described were conducted at the facilities of the (Fundacion IRAUy, Montevideo, Uruguay) and at the (UBAL) of the , Uruguay. Specially, the authors thank Dr. Rosario Durán and Dr. Alejandro Leyva for kindly assisting us in the proteomic experiment. Finnacial support was provided by Fundacion IRAUy; PRONEX 02/2015 (Programa de Apoio a Núcleos de Excelência Pronex/Funcap/CNPq); the Brazilian Research Council?CNPq (grants # 313160/2017‐1 and 438773/2018‐7); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brazil. Instituto de Reproducción Animal Uruguay Unidad de Biotecnología en Animales de Laboratorio Institut Pasteur de Montevideo. -- Corresponding author: A. Moura, A.; Laboratório de Fisiologia e Ciências Ômicas, Departamento de Zootecnia, Universidade Federal do Ceará, Fortaleza, Brazil; email:arlindo.moura@gmail.com -- Menchaca, A.; Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; mail:menchaca.alejo@gmail.com 520 $aABSTRACT. -The present study was conducted to characterize the major proteome of preimplantation (D6) ovine embryos produced in vitro. COCs were aspirated from antral follicles (2–6 mm), matured and fertilized in vitro and cultured until day six. Proteins were ex- tracted separately from three pools of 45 embryos and separately run in SDS-PAGE. Proteins from each pool were individually subjected to in-gel digestion followed by LC-MS/MS. Three ‘raw files’ and protein lists were produced by Pattern Lab software, but only proteins present in all three lists were used for the bioinformatics analyses. There were 2,262 proteins identified in the 6-day-old ovine embryos, including al- bumin, zona pellucida glycoprotein 2, 3 and 4, peptidyl arginine deiminase 6, actin cytoplasmic 1, gamma-actin 1, pyruvate kinase, heat shock protein 90 and protein disulfide isomerase, among others. Major biological processes linked to the sheep embryo proteome were translation, protein transport and protein stabilization, and molecular functions, defined as ATP binding, oxygen carrier activity and oxygen bind- ing. There were 42 enriched functional clusters according to the 2,147 genes (UniProt database). Ten selected clusters with potential association with embryo development included translation, structural constituent of ribosomes, ribosomes, nucleosomes, structural constituent of the cytoskeleton, microtubule-based process, translation initiation factor activity, regulation of translational initiation, cell body and nucleotide biosynthetic process. The most representative KEEG pathways were ribosome, oxida- tive phosphorylation, glutathione metabolism, gap junction, mineral absorption, DNA replication and cGMP-PKG signalling pathway. Analyses of functional clusters clearly showed differences associated with the proteome of preimplantation (D6) sheep em- bryos generated after in vitro fertilization in comparison with in vivo counterparts (Sanchez et al., 2021; https://doi.org/10.1111/rda.13897), confirming that the quality of in vitro derived blastocysts are unlike those produced in vivo. The present study portrays the first comprehensive overview of the proteome of preimplantational ovine embryos grown in vitro. © 2022 Wiley-VCH GmbH. 653 $aEmbryo development 653 $aIn vitro fertilization 653 $aMass spectrometry 653 $aOocyte 653 $aOvine 653 $aPLATAFORMA SALUD ANIMAL 653 $aProteins 700 1 $aGUERREIRO, D. D. 700 1 $aOTÁVIO, K. S. 700 1 $aSANTOS-NETO, P. C. DOS 700 1 $aSOUZA-NEVES, M. 700 1 $aCUADRO, F. 700 1 $aNUÑEZ-OLIVERA, R. 700 1 $aCRISPO, M. 700 1 $aBEZERRA, M. J. B. 700 1 $aSILVA, R. F. 700 1 $aLIMA, L. F. 700 1 $aFIGUEIREDO, J. R. 700 1 $aBUSTAMANTE-FILHO, I. C. 700 1 $aMENCHACA, A. 700 1 $aMOURA, A. A. 773 $tReproduction in Domestic Animals, 2022, Volume 57, Issue 7; pages 784-797. doi: https://doi.org/10.1111/rda.14122
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