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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 : 10 | |
1. | | AREVALO, A.P.; BASIKA, T.; ANCHETA, S.; PERELMUTER, K.; RICCIARDI, A.; BOLLATI-FOGOLÍN, M.; CRISPO, M. 3R's applied to in vivo biological activity of recombinant human erythropoietin assay. Technical Article. Biological Models Research and Technology Journal. 2023, Volume 3, Issue 1, e00012023. http://dx.doi.org/10.4322/2675-9225.00012023 -- OPEN ACCESS. Article history: Submitted date 9 February 2023, Accepted date 19 April 2023. -- Correspondence: Crispo, M.; Institut Pasteur de Montevideo, Laboratory Animals Biotechnology Unit, Montevideo, Uruguay; email:crispo@pasteur.edu.uy --...Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : Internacional - -- |
Biblioteca(s): INIA Las Brujas. |
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2. | | 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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3. | | 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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4. | | CIUFFO, C.; ARÉVALO, A.P.; GARCIA, F.; PERDOMO, T.; CRISPO, M.; GIANNITTI, F.; BUSCHIAZZO, A.; ZARANTONELLI, L. Estudio de la capacidad infectiva, colonización renal e inmunogenicidad de Leptospira interrogans serogrupo Pomona serovar Kennewicki en un modelo murino sub-letal y crónico. [abstract]. Sub-área 4: Patogenia, virulencia y resistencia. In: Anais III Consensos Latinoamericanos em Leptospirose Animal. Nov 30-Dic 02, 2021 (evento online).Tipo: Abstracts/Resúmenes |
Biblioteca(s): INIA Las Brujas. |
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5. | | CIUFFO, C.; ARÉVALO, A.P.; GARCIA, F.; PERDOMO, T.; CRISPO, M.; GIANNITTI, F.; BUSCHIAZZO, A.; ZARANTONELLI, L. Estudio y caracterización de una variante autóctona de Leptospira para su potencial uso como antígeno en vacunas de uso veterinario. In: Jornadas Uruguayas de Buiatría, 49., 2022. Paysandú, Uruguay: Centro Médico Veterinario de Paysandú; Filial de la Sociedad de Medicina Veterinaria del Uruguaya; Sociedad Uruguaya de Buiatría, 9 al 11 Junio 2022. p.192-194. p.192-194.Tipo: Trabajos en Congresos/Conferencias |
Biblioteca(s): INIA Las Brujas. |
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6. | | CRISPO, M.; CHENOUARD, V.; DOS SANTOS-NETO P; TESSON, L.; SOUZA-NEVES, M.; HESLAN, J.M.; CUADRO, F.; ANEGÓN, I.; MENCHACA, A. Generation of a Human Deafness Sheep Model Using the CRISPR/Cas System. Chapter 12. Methods in Molecular Biology, 2022, Volume 2495, Pages 233-258. doi: https://doi.org/10.1007/978-1-0716-2301-5_12Tipo: Capítulo en Libro Técnico-Científico |
Biblioteca(s): INIA La Estanzuela. |
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7. | | ALTIER, N.; ABREO, E.; BEYHAUT, E.; GARAYCOCHEA, S.; TORRES, P.; CERECETTO, V.; MARTÍN, N.; CUITIÑO, M.J.; CRISPO, M.; ARÉVALO, A.P.; REGO, N.; ARROSPIDE, G.; LAGE, M.; SUNDBERG, G. Desarrollo de un biofertilizante microbiano para aumentar la disponibilidad de fósforo en el cultivo de soja. * Sustentabilidad. Revista INIA Uruguay, 2020, no.62, p.95-100. (Revista INIA; 62) *Proyecto: Desarrollo de inoculantes para la movilización de fósforo como insumo en la producción agrícola. INIA/IP/EMPRESAS. Proyecto financiado
por la herramienta Alianzas para la Innovación de la Agencia Nacional de Investigación e...Tipo: Artículos en Revistas Agropecuarias |
Biblioteca(s): INIA Las Brujas. |
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8. | | CABRERA, A.; FRESIA, P.; BERNÁ, L.; SILVEIRA, C.S.; MACÍAS-RIOSECO, M.; AREVALO, A.P.; CRISPO, M.; PRITSCH, O.; RIET-CORREA, F.; GIANNITTI, F.; GIANNITTI, F.; FRANCIA, M.E.; ROBELLO, C. Isolation and molecular characterization of four novel Neospora caninum strains. Genetics, Evolution, and Phylogeny - Short Communication. Parasitology Research, 1 December 2019, Volume 118, Issue 12, Pages 3535-3542. Doi: 10.1007/s00436-019-06474-9 Article history: Received: 11 April 2019 / Accepted: 24 September 2019 / Published online: 7 November 2019.
Funding Sponsor: Agencia Nacional de Investigación e Innovación (ANII).
Funding Text: This project was funded by grant...Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : Internacional - -- |
Biblioteca(s): INIA Las Brujas. |
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9. | | 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. Global proteomic analysis of preimplantational ovine embryos produced in vitro. Reproduction in Domestic Animals, 2022, Volume 57, Issue 7; pages 784-797. doi: https://doi.org/10.1111/rda.14122 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....Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : Internacional - -- |
Biblioteca(s): INIA Las Brujas. |
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10. | | PASSOS, J.R.S.; GUERREIRO, D.D.; KAMILA S. OTÁVIO; DOS SANTOS-NETO, P.C.; SOUZA-NEVES, M.; CUADRO, F.; NUÑEZ?OLIVERA, R.; CRISPO, M.; VASCONCELOS, F.R.; BEZERRA, M.J.B.; SILVA, R.F.; LIMA, L.F.; FIGUEIREDO, J.R.; BUSTAMANTE-FILHO, I.C.; MENCHACA, A.; MOURA, A.A. How in vitro maturation changes the proteome of ovine cumulus-oocyte complexes?. Volume 89, Issue 10, Pages 459 - 470October 2022 Molecular reproduction and development, October 2022, Volume 89, Issue 10, pages 459-470. doi: https://doi.org/10.1002/mrd.23638 Article history: Received: 16 February 2022 | Accepted: 21 July 2022. -- Corresponding author: Moura, A.A.; Laboratory of Animal Physiology, Department of Animal Science, Federal University of Ceará, Fortaleza, Brazil;...Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : Internacional - -- |
Biblioteca(s): INIA La Estanzuela. |
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Registros recuperados : 10 | |
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