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| Acceso al texto completo restringido a Biblioteca INIA Las Brujas. Por información adicional contacte bibliolb@inia.org.uy. |
Registro completo
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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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Registros recuperados : 28 | |
1. | | MENCHACA, A. Assisted Reproductive Technologies (ART) and genome editing to support a sustainable livestock. Thematic Section: 36th Annual Meeting of the Brazilian Embryo Technology Society (SBTE). Animal Reproduction, 2023, Volume 20, Issue 2, e20230074. http://dx.doi.org/10.1590/1984-3143-AR2023-0074 -- OPEN ACCESS. Article history: Received May 26, 2023; Accepted July 21, 2023. -- Document type: Article Gold Open Access, Green Open Access. -- Correspondence: Menchaca, A.; Plataforma de Salud Animal, Instituto Nacional de Investigación...Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : Internacional - -- |
Biblioteca(s): INIA Las Brujas. |
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3. | | MENCHACA, A. Sustainable food production: The contribution of genome editing in livestock. Sustainability (Switzerland), June 2021, volume 13, Issue 122, article 6788. OPEN ACCESS. Doi: https://doi.org/10.3390/su13126788 Article history: Received 16 March 2021; Revised 31 May 2021; Accepted 1 June 2021; Published 16 June 2021.
Academic Editor: Gerhart U. Ryffel.
This article belongs to the Special Issue Genome Editing in Animal System to Support...Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : Internacional - -- |
Biblioteca(s): INIA Las Brujas. |
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4. | | BÓ, G.A.; MENCHACA, A. Prohibition of hormones in animal reproduction: what to expect and what to do?. Thematic Section: 36th Annual Meeting of the Brazilian Embryo Technology Society (SBTE). Animal Reproduction, 2023, Volume 20, Issue 2, e20230067. https://doi.org/10.1590/1984-3143-AR2023-0067 -- OPEN ACCESS. Article history: Received May 15, 2023; Accepted: July 18, 2023. -- Financial support: GAB received funding for this research from Fondo Nacional de Ciencia y Tecnología (FONCYT PICT 2017-4550), Instituto de Investigación Universidad...Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : Internacional - -- |
Biblioteca(s): INIA Las Brujas. |
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8. | | GARCÍA PINTOS, C.; RIET-CORREA, F.; MENCHACA, A. Effect of foot-and-mouth disease vaccine on pregnancy failure in beef cows. Frontiers in Veterinary Science, November 2021, volume 8, Article number 761304. Open Access. doi: https://doi.org/10.3389/fvets.2021.761304 Article history: Received 19 August 2021; Accepted 18 October 2021; Published 12 November 2021.
Corresponding author: Menchaca, A.; Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay;...Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : Internacional - -- |
Biblioteca(s): INIA Las Brujas. |
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12. | | KALDS, P.; CRISPO, M.; TESSON, L.; ANEGÓN, I.; CHEN KEY, Y.; WANG, X.; MENCHACA, A. Generation of Double-Muscled Sheep and Goats by CRISPR /Cas9-Mediated Knockout of the Myostatin Gene. Chapter 16. Methods in Molecular Biology, 2022, Volume 2495, Pages 295-323. Doi: https://doi.org/10.1007/978-1-0716-2301-5_16Tipo: Capítulo en Libro Técnico-Científico |
Biblioteca(s): INIA La Estanzuela. |
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17. | | NOVAS, R.; BASIKA, T.; WILLIAMSON, M. E.; FRESIA, P.; MENCHACA, A.; SCOTT, M. J. Identification and functional analysis of Cochliomyia hominivorax U6 gene promoters. Insect Molecular Biology, 2023. Early View. https://doi.org/10.1111/imb.12875 -- OPEN ACCESS. Article history: Received 9 March 2023, Accepted 4 August 2023, First published 21 September 2023. -- Scott, M.J.; Department of Entomology and Plant Pathology, North Carolina State University, Campus Box 7613, Raleigh, NC, United...Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : Internacional - -- |
Biblioteca(s): INIA Las Brujas. |
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19. | | DOS SANTOS-NETO, P.C.; CUADRO, F.; SOUZA-NEVES, M.; CRISPO, M.; MENCHACA, A. Refinements in embryo manipulation applied to CRISPR technology in livestock. Theriogenology. 2023, Volume 208, Pages 142-148. https://doi.org/10.1016/j.theriogenology.2023.05.028 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,...Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : Internacional - -- |
Biblioteca(s): INIA Las Brujas. |
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20. | | CUADRO, F.; DE BRUN, V.; BROCHADO, C.; SOUZA, M.; GARCÍA PINTOS, C.; MENEZES, C.; GASTAL, G.D.A.; MENCHACA, A. Gene expression patterns in uterus and oviduct during the preovulatory period in ewes. In: 19th International Congress on Animal Reproduction (ICAR), Bologna (Italy), 26-30 June 2022. p.185.Biblioteca(s): INIA La Estanzuela. |
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Registros recuperados : 28 | |
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