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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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Registro completo
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Biblioteca (s) : |
INIA Las Brujas. |
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Fecha : |
13/07/2023 |
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Actualizado : |
13/07/2023 |
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Tipo de producción científica : |
Artículos en Revistas Indexadas Internacionales |
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Autor : |
DOS SANTOS-NETO, P.C.; CUADRO, F.; SOUZA-NEVES, M.; CRISPO, M.; MENCHACA, A. |
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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. |
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Título : |
Refinements in embryo manipulation applied to CRISPR technology in livestock. |
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Fecha de publicación : |
2023 |
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Fuente / Imprenta : |
Theriogenology. 2023, Volume 208, Pages 142-148. https://doi.org/10.1016/j.theriogenology.2023.05.028 |
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ISSN : |
0093-691X |
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DOI : |
10.1016/j.theriogenology.2023.05.028 |
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Idioma : |
Inglés |
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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. -- |
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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 |
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Palabras claves : |
Early embryo transfer; Gene editing; In vitro embryo production; Minimum volume vitrification. |
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Asunto categoría : |
L10 Genética y mejoramiento animal |
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Marc : |
LEADER 04268naa a2200253 a 4500
001 1064251
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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 original : |
INIA Las Brujas (LB) |
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Registro completo
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Biblioteca (s) : |
INIA Las Brujas. |
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Fecha actual : |
12/08/2015 |
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Actualizado : |
12/06/2018 |
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Tipo de producción científica : |
Documentos |
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Autor : |
BRANDA, A.; FEDERICI, M.; BRIANO, C.; ROMERO, A.; LLAMBÍ, S.; DUTRA, F.; DALLA RIZZA, M. |
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Afiliación : |
ANDREA BRANDA SICA, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; MARIA TERESA FEDERICI RODRIGUEZ, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; C. BRIANO, MGAP/ DILAVE (División de Laboratorios de Veterinarios); A. ROMERO, MGAP/ DILAVE (División de Laboratorios de Veterinarios); S. LLAMBÍ, Universidad de la República (UdelaR)/ Facultad de Veterinaria; F. DUTRA, MGAP/ DILAVE (División de Laboratorios de Veterinarios); MARCO DALLA RIZZA VILARO, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay. |
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Título : |
Avances en la identificación de portadores de la deficiencia en la adhesión leucocitaria bovina en raza Holando. |
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Fecha de publicación : |
2014 |
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Fuente / Imprenta : |
In: JORNADA TÉCNICA, VIII JORNADA DE AGROBIOTECNOLOGÍA. INIA LAS BRUJAS, 30 DE OCTUBRE DE 2014. UNIDAD DE BIOTECNOLOGÍA. Montevideo (Uruguay): INIA, 2014. |
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Páginas : |
7-9 |
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Serie : |
(Actividades de Difusión; 741) |
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ISSN : |
1688-9258 |
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Idioma : |
Español |
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Contenido : |
La deficiencia en la adhesión leucocitaria bovina (BLAD) representa un problema a nivel de los tambos por generar inmunodepresión que podría ser la causa predisponente de muchas de las patologías infecciosas reconocidas en terneros. Los animales afectados mueren a causa de la extrema susceptibilidad a las infecciones, causada por la incapacidad de glóbulos blancos (leucocitos) para pasar al espacio extravascular en el tejido infectado. La deficiencia en la adhesión leucocitaria bovina (BLAD) representa un problema a nivel de los tambos por generar inmunodepresión que podría ser la causa predisponente de muchas de las patologías infecciosas reconocidas en terneros. Los animales afectados mueren a causa de la extrema susceptibilidad a las infecciones, causada por la incapacidad de glóbulos blancos
(leucocitos) para pasar al espacio extravascular en el tejido infectado. |
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Palabras claves : |
BANCO DE ADN GENÓMICO ANIMAL; BLAD. |
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Thesagro : |
ENFERMEDADES DE LOS ANIMALES; GANADO DE LECHE; HOLANDO; TECNICAS DE DIAGNOSIS. |
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Asunto categoría : |
L73 Enfermedades de los animales |
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URL : |
http://www.ainfo.inia.uy/digital/bitstream/item/10172/1/sad-741-p.7-9.pdf
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Marc : |
LEADER 01850naa a2200301 a 4500
001 1053214
005 2018-06-12
008 2014 bl uuuu u00u1 u #d
022 $a1688-9258
100 1 $aBRANDA, A.
245 $aAvances en la identificación de portadores de la deficiencia en la adhesión leucocitaria bovina en raza Holando.
260 $c2014
300 $a7-9
490 $a(Actividades de Difusión; 741)
520 $aLa deficiencia en la adhesión leucocitaria bovina (BLAD) representa un problema a nivel de los tambos por generar inmunodepresión que podría ser la causa predisponente de muchas de las patologías infecciosas reconocidas en terneros. Los animales afectados mueren a causa de la extrema susceptibilidad a las infecciones, causada por la incapacidad de glóbulos blancos (leucocitos) para pasar al espacio extravascular en el tejido infectado. La deficiencia en la adhesión leucocitaria bovina (BLAD) representa un problema a nivel de los tambos por generar inmunodepresión que podría ser la causa predisponente de muchas de las patologías infecciosas reconocidas en terneros. Los animales afectados mueren a causa de la extrema susceptibilidad a las infecciones, causada por la incapacidad de glóbulos blancos (leucocitos) para pasar al espacio extravascular en el tejido infectado.
650 $aENFERMEDADES DE LOS ANIMALES
650 $aGANADO DE LECHE
650 $aHOLANDO
650 $aTECNICAS DE DIAGNOSIS
653 $aBANCO DE ADN GENÓMICO ANIMAL
653 $aBLAD
700 1 $aFEDERICI, M.
700 1 $aBRIANO, C.
700 1 $aROMERO, A.
700 1 $aLLAMBÍ, S.
700 1 $aDUTRA, F.
700 1 $aDALLA RIZZA, M.
773 $tIn: JORNADA TÉCNICA, VIII JORNADA DE AGROBIOTECNOLOGÍA. INIA LAS BRUJAS, 30 DE OCTUBRE DE 2014. UNIDAD DE BIOTECNOLOGÍA. Montevideo (Uruguay): INIA, 2014.
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