dc.contributor.author | Navarro Serna, Sergio | |
dc.contributor.author | Piñeiro Silva, Celia | |
dc.contributor.author | Fernández Martín, Irene | |
dc.contributor.author | Dehesa Etxebeste, Martxel Pedro | |
dc.contributor.author | López de Munain Arregui, Adolfo José | |
dc.contributor.author | Gadea, Joaquín | |
dc.date.accessioned | 2024-05-15T17:30:15Z | |
dc.date.available | 2024-05-15T17:30:15Z | |
dc.date.issued | 2024-04 | |
dc.identifier.citation | Theriogenology 218 : 111-118 (2024) | es_ES |
dc.identifier.issn | 0093-691X | |
dc.identifier.issn | 1879-3231 | |
dc.identifier.uri | http://hdl.handle.net/10810/67971 | |
dc.description.abstract | Genetically modified pigs play a critical role in mimicking human diseases, xenotransplantation, and the development of pigs resistant to viral diseases. The use of programmable endonucleases, including the CRISPR/Cas9 system, has revolutionized the generation of genetically modified pigs. This study evaluates the efficiency of electroporation of oocytes prior to fertilization in generating edited gene embryos for different models. For single gene editing, phospholipase C zeta (PLC ζ) and fused in sarcoma (FUS) genes were used, and the concentration of sgRNA and Cas9 complexes was optimized. The results showed that increasing the concentration resulted in higher mutation rates without affecting the blastocyst rate. Electroporation produced double knockouts for the TPC1/TPC2 genes with high efficiency (79 %). In addition, resistance to viral diseases such as PRRS and swine influenza was achieved by electroporation, allowing the generation of double knockout embryo pigs (63 %). The study also demonstrated the potential for multiple gene editing in a single step using electroporation, which is relevant for xenotransplantation. The technique resulted in the simultaneous mutation of 5 genes (GGTA1, B4GALNT2, pseudo B4GALNT2, CMAH and GHR). Overall, electroporation proved to be an efficient and versatile method to generate genetically modified embryonic pigs, offering significant advances in biomedical and agricultural research, xenotransplantation, and disease resistance. Electroporation led to the processing of numerous oocytes in a single session using less expensive equipment. We confirmed the generation of gene-edited porcine embryos for single, double, or quintuple genes simultaneously without altering embryo development to the blastocyst stage. The results provide valuable insights into the optimization of gene editing protocols for different models, opening new avenues for research and applications in this field. | es_ES |
dc.description.sponsorship | This research was funded by Fundación Séneca 22065/PI/22, Instituto de Salud Carlos III Proyectos de Desarrollo Tecnológico AES 2019 (DTS19/00061), Spanish Ministry of Science, Innovation and Universities: MICIN PID2020-113366RB-I00, MCIN/AEI/10.13039/501100011033/, FEDER Una manera de hacer Europa; Universidad de Murcia predoctoral fellowship R-496/2022. | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | Elsevier | es_ES |
dc.relation | info:eu-repo/grantAgreement/MICINN/PID2020-113366RB-I00 | es_ES |
dc.rights | info:eu-repo/semantics/openAccess | es_ES |
dc.rights.uri | http://creativecommons.org/licenses/by/3.0/es/ | * |
dc.subject | phospholipase C zeta (PLC ζ) | es_ES |
dc.subject | fused in sarcoma (FUS) | es_ES |
dc.subject | two pore channels (TPC1 and TPC2) | es_ES |
dc.subject | CD163 | es_ES |
dc.subject | CRISPR/Cas9 | es_ES |
dc.subject | gene editing | es_ES |
dc.subject | Knockout porcine embryos | es_ES |
dc.subject | biomedical research | es_ES |
dc.subject | xenotransplantation | es_ES |
dc.subject | disease resistance | es_ES |
dc.title | Oocyte electroporation prior to in vitro fertilization is an efficient method to generate single, double, and multiple knockout porcine embryos of interest in biomedicine and animal production | es_ES |
dc.type | info:eu-repo/semantics/article | es_ES |
dc.rights.holder | © 2024 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/) | es_ES |
dc.rights.holder | Atribución 3.0 España | * |
dc.relation.publisherversion | https://www.sciencedirect.com/science/article/pii/S0093691X24000578 | es_ES |
dc.identifier.doi | 10.1016/j.theriogenology.2024.01.040 | |
dc.departamentoes | Neurociencias | es_ES |
dc.departamentoeu | Neurozientziak | es_ES |