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Biblioteca (s) : |
INIA Las Brujas. |
Fecha : |
13/07/2023 |
Actualizado : |
13/07/2023 |
Tipo de producción científica : |
Artículos en Revistas Indexadas Internacionales |
Autor : |
DOS SANTOS-NETO, P.C.; CUADRO, F.; SOUZA-NEVES, M.; CRISPO, M.; MENCHACA, A. |
Afiliación : |
P.C. DOS SANTOS-NETO, Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; F. CUADRO, Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; M. SOUZA-NEVES, Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; Unidad de Biotecnología en Animales de Laboratorio, Institut Pasteur de Montevideo, Uruguay; M. CRISPO, Unidad de Biotecnología en Animales de Laboratorio, Institut Pasteur de Montevideo, Uruguay; JOSE ALEJO MENCHACA BARBEITO, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay. |
Título : |
Refinements in embryo manipulation applied to CRISPR technology in livestock. |
Fecha de publicación : |
2023 |
Fuente / Imprenta : |
Theriogenology. 2023, Volume 208, Pages 142-148. https://doi.org/10.1016/j.theriogenology.2023.05.028 |
ISSN : |
0093-691X |
DOI : |
10.1016/j.theriogenology.2023.05.028 |
Idioma : |
Inglés |
Notas : |
Article history: Received 14 April 2023; Received in revised form 29 May 2023; Accepted 29 May 2023; Available online 9 June 2023. -- Correspondence author: Menchaca, A.; Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; email:menchaca.alejo@gmail.com -- FUNDING: This study was financially supported by Fundación IRAUy, FOCEM (MERCOSUR Structural Convergence Fund), COF 03/11, and the Uruguayan National Research Agency (ANII, Uruguay). PCdSN received a scholarship from the National Council for Scientific Technological Development (CNPq, Brazil). AM, MC, and PCdSN are fellows of Sistema Nacional de Investigadores (SNI, ANII) of Uruguay and PEDECIBA. -- |
Contenido : |
The implementation of CRISPR technology in large animals requires further improvements in embryo manipulation and transfer to be applied with commercial purposes. In this study we report (a) developmental competence of CRISPR/Cas microinjected zygotes subjected to in vitro culture in large scale programs in sheep; (b) pregnancy outcomes after early-stage (2-8-cell) embryo transfer into the oviduct or the uterine horn; and (c) embryo survival and birth rate after vitrification/warming of CRISPR/Cas microinjected zygotes. Experiment 1 consisted of a retrospective analysis to evaluate embryo developmental rate of in vitro produced zygotes subjected to CRISPR/Cas microinjection (n = 7,819) compared with a subset of non-microinjected zygotes (n = 701). Development rates to blastocyst on Day 6 were 20.0% for microinjected zygotes and 44.9% for non-injected zygotes (P < 0.05). In Experiment 2, CRISPR/Cas microinjected zygotes were transferred on Day 2 after in vitro fertilization (2-8 cell embryos) into the oviductal ampulla (n = 262) or into the uterine horn (n = 276) in synchronized recipient ewes at prefixed time (i.e., approximately two days after ovulation). Pregnant/transferred recipients (24.0% vs. 25.0%), embryo survival/transferred embryos (6.9% vs. 6.2%), and born lambs/pregnant embryos (72.2% vs. 100.0%) did not differ significantly in the two groups. In Experiment 3, CRISPR/Cas microinjected zygotes were maintained under in vitro culture until blastocyst stage (Day 6), and subjected to vitrification/warming via the Cryotop method (n = 474), while a subset of embryos were left fresh as control group (n = 75). Embryos were transferred into the uterine horn of recipient females at prefixed time 8.5 days after the estrous synchronization treatment (i.e., approximately six days after ovulation). Pregnancy rate (30.8% vs. 48.0%), embryo survival rate (14.8% vs. 21.3%), and birth rate (85.7% vs. 75.0%) were not different (P[dbnd]NS) between vitrified and fresh embryos, respectively. In conclusion, the current study in sheep embryos reports (a) suitable developmental rate after CRISPR/Cas microinjection (i.e., 20%), even though it was lower than non-microinjected zygotes; (b) similar outcomes when Day 2-embryos were placed into the uterine horn instead of the oviduct, avoiding both time-consuming and invasive oviduct manipulation, and extended in vitro culture during one week; (c) promising pregnancy and birth rates obtained with vitrification of CRISPR/Cas microinjected embryos. This knowledge on in vitro embryo development, timing of embryo transfer, and cryopreservation of CRISPR/Cas microinjected zygotes have practical implications for the implementation of genome editing technology in large animals. © 2023 Elsevier Inc. MenosThe implementation of CRISPR technology in large animals requires further improvements in embryo manipulation and transfer to be applied with commercial purposes. In this study we report (a) developmental competence of CRISPR/Cas microinjected zygotes subjected to in vitro culture in large scale programs in sheep; (b) pregnancy outcomes after early-stage (2-8-cell) embryo transfer into the oviduct or the uterine horn; and (c) embryo survival and birth rate after vitrification/warming of CRISPR/Cas microinjected zygotes. Experiment 1 consisted of a retrospective analysis to evaluate embryo developmental rate of in vitro produced zygotes subjected to CRISPR/Cas microinjection (n = 7,819) compared with a subset of non-microinjected zygotes (n = 701). Development rates to blastocyst on Day 6 were 20.0% for microinjected zygotes and 44.9% for non-injected zygotes (P < 0.05). In Experiment 2, CRISPR/Cas microinjected zygotes were transferred on Day 2 after in vitro fertilization (2-8 cell embryos) into the oviductal ampulla (n = 262) or into the uterine horn (n = 276) in synchronized recipient ewes at prefixed time (i.e., approximately two days after ovulation). Pregnant/transferred recipients (24.0% vs. 25.0%), embryo survival/transferred embryos (6.9% vs. 6.2%), and born lambs/pregnant embryos (72.2% vs. 100.0%) did not differ significantly in the two groups. In Experiment 3, CRISPR/Cas microinjected zygotes were maintained under in vitro culture until blastocyst stage (Day 6), ... Presentar Todo |
Palabras claves : |
Early embryo transfer; Gene editing; In vitro embryo production; Minimum volume vitrification. |
Asunto categoría : |
L10 Genética y mejoramiento animal |
Marc : |
LEADER 04268naa a2200253 a 4500 001 1064251 005 2023-07-13 008 2023 bl uuuu u00u1 u #d 022 $a0093-691X 024 7 $a10.1016/j.theriogenology.2023.05.028$2DOI 100 1 $aDOS SANTOS-NETO, P.C. 245 $aRefinements in embryo manipulation applied to CRISPR technology in livestock.$h[electronic resource] 260 $c2023 500 $aArticle history: Received 14 April 2023; Received in revised form 29 May 2023; Accepted 29 May 2023; Available online 9 June 2023. -- Correspondence author: Menchaca, A.; Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay; email:menchaca.alejo@gmail.com -- FUNDING: This study was financially supported by Fundación IRAUy, FOCEM (MERCOSUR Structural Convergence Fund), COF 03/11, and the Uruguayan National Research Agency (ANII, Uruguay). PCdSN received a scholarship from the National Council for Scientific Technological Development (CNPq, Brazil). AM, MC, and PCdSN are fellows of Sistema Nacional de Investigadores (SNI, ANII) of Uruguay and PEDECIBA. -- 520 $aThe implementation of CRISPR technology in large animals requires further improvements in embryo manipulation and transfer to be applied with commercial purposes. In this study we report (a) developmental competence of CRISPR/Cas microinjected zygotes subjected to in vitro culture in large scale programs in sheep; (b) pregnancy outcomes after early-stage (2-8-cell) embryo transfer into the oviduct or the uterine horn; and (c) embryo survival and birth rate after vitrification/warming of CRISPR/Cas microinjected zygotes. Experiment 1 consisted of a retrospective analysis to evaluate embryo developmental rate of in vitro produced zygotes subjected to CRISPR/Cas microinjection (n = 7,819) compared with a subset of non-microinjected zygotes (n = 701). Development rates to blastocyst on Day 6 were 20.0% for microinjected zygotes and 44.9% for non-injected zygotes (P < 0.05). In Experiment 2, CRISPR/Cas microinjected zygotes were transferred on Day 2 after in vitro fertilization (2-8 cell embryos) into the oviductal ampulla (n = 262) or into the uterine horn (n = 276) in synchronized recipient ewes at prefixed time (i.e., approximately two days after ovulation). Pregnant/transferred recipients (24.0% vs. 25.0%), embryo survival/transferred embryos (6.9% vs. 6.2%), and born lambs/pregnant embryos (72.2% vs. 100.0%) did not differ significantly in the two groups. In Experiment 3, CRISPR/Cas microinjected zygotes were maintained under in vitro culture until blastocyst stage (Day 6), and subjected to vitrification/warming via the Cryotop method (n = 474), while a subset of embryos were left fresh as control group (n = 75). Embryos were transferred into the uterine horn of recipient females at prefixed time 8.5 days after the estrous synchronization treatment (i.e., approximately six days after ovulation). Pregnancy rate (30.8% vs. 48.0%), embryo survival rate (14.8% vs. 21.3%), and birth rate (85.7% vs. 75.0%) were not different (P[dbnd]NS) between vitrified and fresh embryos, respectively. In conclusion, the current study in sheep embryos reports (a) suitable developmental rate after CRISPR/Cas microinjection (i.e., 20%), even though it was lower than non-microinjected zygotes; (b) similar outcomes when Day 2-embryos were placed into the uterine horn instead of the oviduct, avoiding both time-consuming and invasive oviduct manipulation, and extended in vitro culture during one week; (c) promising pregnancy and birth rates obtained with vitrification of CRISPR/Cas microinjected embryos. This knowledge on in vitro embryo development, timing of embryo transfer, and cryopreservation of CRISPR/Cas microinjected zygotes have practical implications for the implementation of genome editing technology in large animals. © 2023 Elsevier Inc. 653 $aEarly embryo transfer 653 $aGene editing 653 $aIn vitro embryo production 653 $aMinimum volume vitrification 700 1 $aCUADRO, F. 700 1 $aSOUZA-NEVES, M. 700 1 $aCRISPO, M. 700 1 $aMENCHACA, A. 773 $tTheriogenology. 2023, Volume 208, Pages 142-148. https://doi.org/10.1016/j.theriogenology.2023.05.028
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Registro completo
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Biblioteca (s) : |
INIA Las Brujas. |
Fecha actual : |
20/06/2015 |
Actualizado : |
20/06/2015 |
Tipo de producción científica : |
Informes Agroclimáticos |
Autor : |
GIMENEZ, A.; CASTAÑO, J.; FUREST, J.; CAL, A.; TISCORNIA, G.; SCHIAVI, C. |
Afiliación : |
AGUSTIN EDUARDO GIMENEZ FUREST, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; JOSE PEDRO CASTAÑO SANCHEZ, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; JOSE MARIA FUREST CROCCO, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; ADRIAN TABARE CAL ALVAREZ, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; GUADALUPE TISCORNIA TOSAR, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; CARLOS IGNACIO SCHIAVI RAMPELBERG, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay. |
Título : |
Informe Agroclimático 2013 - Situación a Abril. |
Fecha de publicación : |
2013 |
Fuente / Imprenta : |
Montevideo (Uruguay): INIA, 2013. |
Páginas : |
4 p. |
Idioma : |
Español |
Palabras claves : |
AGROCLIMA; AGROCLIMATOLOGÍA; BOLETIN AGROCLIMÁTICO; CARACTERIZACIÓN AGROCLIMÁTICA; DIRECCION VIENTO; ESTACIONES AGROMETEOROLOGICAS; ESTACIONES AUTOMATICAS; ESTACIONES INIA; ESTADO DEL TIEMPO; ESTRÉS HÍDRICO; GRAFICAS AGROCLIMATICOS; GRAS; HELIOFANOGRAFO; INFORMACION SATELITAL; INUNDACIONES; LLUVIAS DIARIAS; MAXIMA; MEDIA; MINIMA; PANEL SOLAR; PERSPECTIVAS CLIMATICAS; PLUVIOMETRO; PRECIPITACION NACIONAL; PREVENCION HELADAS; PRONOSTICO; SENSOR; SIMETRICO; TANQUE A; TERMOCUPLAS; TERMOHIDROGRAFO; VARIABLES AGROCLIMATICAS; VELETA. |
Thesagro : |
AGROCLIMATOLOGIA; CAMBIO CLIMATICO; CLIMA; CLIMATOLOGIA; ESTACIONES METEOROLOGICAS; ESTRES HIDRICO; EVAPORACION; EVAPOTRANSPIRACION; HUMEDAD; HUMEDAD RELATIVA; LLUVIA; METEOROLOGIA; PERSPECTIVAS; PLUVIOMETROS; PRONOSTICO DEL TIEMPO; SENSORES; SISTEMAS; SISTEMAS DE INFORMACION; SUELO; TEMPERATURA; TERMOMETROS. |
Asunto categoría : |
P40 Meteorología y climatología |
URL : |
http://www.ainfo.inia.uy/digital/bitstream/item/4722/1/Inf.Agr.-abril-2013.pdf
http://www.inia.uy/Publicaciones/Paginas/Informe-agroclimatico-2013-Situacion-Abril.aspx
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Marc : |
LEADER 02091nam a2200805 a 4500 001 1052878 005 2015-06-20 008 2013 bl uuuu u0uu1 u #d 100 1 $aGIMENEZ, A. 245 $aInforme Agroclimático 2013 - Situación a Abril.$h[electronic resource] 260 $aMontevideo (Uruguay): INIA$c2013 300 $a4 p. 650 $aAGROCLIMATOLOGIA 650 $aCAMBIO CLIMATICO 650 $aCLIMA 650 $aCLIMATOLOGIA 650 $aESTACIONES METEOROLOGICAS 650 $aESTRES HIDRICO 650 $aEVAPORACION 650 $aEVAPOTRANSPIRACION 650 $aHUMEDAD 650 $aHUMEDAD RELATIVA 650 $aLLUVIA 650 $aMETEOROLOGIA 650 $aPERSPECTIVAS 650 $aPLUVIOMETROS 650 $aPRONOSTICO DEL TIEMPO 650 $aSENSORES 650 $aSISTEMAS 650 $aSISTEMAS DE INFORMACION 650 $aSUELO 650 $aTEMPERATURA 650 $aTERMOMETROS 653 $aAGROCLIMA 653 $aAGROCLIMATOLOGÍA 653 $aBOLETIN AGROCLIMÁTICO 653 $aCARACTERIZACIÓN AGROCLIMÁTICA 653 $aDIRECCION VIENTO 653 $aESTACIONES AGROMETEOROLOGICAS 653 $aESTACIONES AUTOMATICAS 653 $aESTACIONES INIA 653 $aESTADO DEL TIEMPO 653 $aESTRÉS HÍDRICO 653 $aGRAFICAS AGROCLIMATICOS 653 $aGRAS 653 $aHELIOFANOGRAFO 653 $aINFORMACION SATELITAL 653 $aINUNDACIONES 653 $aLLUVIAS DIARIAS 653 $aMAXIMA 653 $aMEDIA 653 $aMINIMA 653 $aPANEL SOLAR 653 $aPERSPECTIVAS CLIMATICAS 653 $aPLUVIOMETRO 653 $aPRECIPITACION NACIONAL 653 $aPREVENCION HELADAS 653 $aPRONOSTICO 653 $aSENSOR 653 $aSIMETRICO 653 $aTANQUE A 653 $aTERMOCUPLAS 653 $aTERMOHIDROGRAFO 653 $aVARIABLES AGROCLIMATICAS 653 $aVELETA 700 1 $aCASTAÑO, J. 700 1 $aFUREST, J. 700 1 $aCAL, A. 700 1 $aTISCORNIA, G. 700 1 $aSCHIAVI, C.
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