Gene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesis
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional do INPA |
Texto Completo: | https://repositorio.inpa.gov.br/handle/1/15219 |
Resumo: | Microinjection is commonly performed to achieve fish transgenesis; however, due to difficulties associated with this technique, new strategies are being developed. Here we evaluate the potential of lentiviral particles to genetically modify Nile tilapia cells to achieve transgenesis using three different approaches: spermatogonial stem cell (SSC) genetic modification and transplantation (SC), in vivo transduction of gametes (GT), and fertilised egg transduction (ET). The SC protocol using larvae generates animals with sustained production of modified sperm (80% of animals with 77% maximum sperm fluorescence [MSF]), but is a time-consuming protocol (sexual maturity in Nile tilapia is achieved at 6 months of age). GT is a faster technique, but the modified gamete production is temporary (70% of animals with 52% MSF). ET is an easier way to obtain mosaic transgenic animals compared to microinjection of eggs, but non-site-directed integration in the fish genome can be a problem. In this study, PI3Kc2α gene disruption impaired development during the embryo stage and caused premature death. The manipulator should choose a technique based on the time available for transgenic obtainment and if this generation is required to be continuous or not. © The Author(s) 2017. |
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Tonelli, Fernanda MPdos Santos NassifLacerda, Samyra MariaProcópio, Marcela SantosLemos, Breno Luiz SalesFrança, Luiz Renato deResende, Rodrigo Ribeiro2020-05-07T14:14:47Z2020-05-07T14:14:47Z2017https://repositorio.inpa.gov.br/handle/1/1521910.1038/srep44317Microinjection is commonly performed to achieve fish transgenesis; however, due to difficulties associated with this technique, new strategies are being developed. Here we evaluate the potential of lentiviral particles to genetically modify Nile tilapia cells to achieve transgenesis using three different approaches: spermatogonial stem cell (SSC) genetic modification and transplantation (SC), in vivo transduction of gametes (GT), and fertilised egg transduction (ET). The SC protocol using larvae generates animals with sustained production of modified sperm (80% of animals with 77% maximum sperm fluorescence [MSF]), but is a time-consuming protocol (sexual maturity in Nile tilapia is achieved at 6 months of age). GT is a faster technique, but the modified gamete production is temporary (70% of animals with 52% MSF). ET is an easier way to obtain mosaic transgenic animals compared to microinjection of eggs, but non-site-directed integration in the fish genome can be a problem. In this study, PI3Kc2α gene disruption impaired development during the embryo stage and caused premature death. The manipulator should choose a technique based on the time available for transgenic obtainment and if this generation is required to be continuous or not. © The Author(s) 2017.Volume 7Attribution-NonCommercial-NoDerivs 3.0 Brazilhttp://creativecommons.org/licenses/by-nc-nd/3.0/br/info:eu-repo/semantics/openAccessPhosphatidylinositol 3 KinaseAdult Stem CellAngiogenesisAnimalsCichlidCytologyDeficiencyFemaleGene Expression RegulationTransduction, GeneticGeneticsGerm CellGrowth, Development And AgingLarvaMaleMetabolismMicro-injectionMutationEmbryo, NonmammalianProceduresTransgenic AnimalsTransplantationVascularizationZygoteAdult Germline Stem CellsAnimalAnimal, Genetically ModifiedCichlidsEmbryo, NonmammalianFemaleGene Expression Regulation, DevelopmentalGerm CellsLarvaMaleMicroinjectionsMutationNeovascularization, PhysiologicPhosphatidylinositol 3-kinasesTransduction, GeneticZygoteGene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesisinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleScientific Reportsengreponame:Repositório Institucional do INPAinstname:Instituto Nacional de Pesquisas da Amazônia (INPA)instacron:INPAORIGINALartigo-inpa.pdfapplication/pdf3681839https://repositorio.inpa.gov.br/bitstream/1/15219/1/artigo-inpa.pdf9140b65f8169fdfa8706c26c5a8c4fcdMD51CC-LICENSElicense_rdfapplication/octet-stream914https://repositorio.inpa.gov.br/bitstream/1/15219/2/license_rdf4d2950bda3d176f570a9f8b328dfbbefMD521/152192020-07-14 10:59:10.806oai:repositorio:1/15219Repositório de PublicaçõesPUBhttps://repositorio.inpa.gov.br/oai/requestopendoar:2020-07-14T14:59:10Repositório Institucional do INPA - Instituto Nacional de Pesquisas da Amazônia (INPA)false |
dc.title.en.fl_str_mv |
Gene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesis |
title |
Gene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesis |
spellingShingle |
Gene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesis Tonelli, Fernanda MP Phosphatidylinositol 3 Kinase Adult Stem Cell Angiogenesis Animals Cichlid Cytology Deficiency Female Gene Expression Regulation Transduction, Genetic Genetics Germ Cell Growth, Development And Aging Larva Male Metabolism Micro-injection Mutation Embryo, Nonmammalian Procedures Transgenic Animals Transplantation Vascularization Zygote Adult Germline Stem Cells Animal Animal, Genetically Modified Cichlids Embryo, Nonmammalian Female Gene Expression Regulation, Developmental Germ Cells Larva Male Microinjections Mutation Neovascularization, Physiologic Phosphatidylinositol 3-kinases Transduction, Genetic Zygote |
title_short |
Gene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesis |
title_full |
Gene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesis |
title_fullStr |
Gene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesis |
title_full_unstemmed |
Gene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesis |
title_sort |
Gene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesis |
author |
Tonelli, Fernanda MP |
author_facet |
Tonelli, Fernanda MP dos Santos NassifLacerda, Samyra Maria Procópio, Marcela Santos Lemos, Breno Luiz Sales França, Luiz Renato de Resende, Rodrigo Ribeiro |
author_role |
author |
author2 |
dos Santos NassifLacerda, Samyra Maria Procópio, Marcela Santos Lemos, Breno Luiz Sales França, Luiz Renato de Resende, Rodrigo Ribeiro |
author2_role |
author author author author author |
dc.contributor.author.fl_str_mv |
Tonelli, Fernanda MP dos Santos NassifLacerda, Samyra Maria Procópio, Marcela Santos Lemos, Breno Luiz Sales França, Luiz Renato de Resende, Rodrigo Ribeiro |
dc.subject.eng.fl_str_mv |
Phosphatidylinositol 3 Kinase Adult Stem Cell Angiogenesis Animals Cichlid Cytology Deficiency Female Gene Expression Regulation Transduction, Genetic Genetics Germ Cell Growth, Development And Aging Larva Male Metabolism Micro-injection Mutation Embryo, Nonmammalian Procedures Transgenic Animals Transplantation Vascularization Zygote Adult Germline Stem Cells Animal Animal, Genetically Modified Cichlids Embryo, Nonmammalian Female Gene Expression Regulation, Developmental Germ Cells Larva Male Microinjections Mutation Neovascularization, Physiologic Phosphatidylinositol 3-kinases Transduction, Genetic Zygote |
topic |
Phosphatidylinositol 3 Kinase Adult Stem Cell Angiogenesis Animals Cichlid Cytology Deficiency Female Gene Expression Regulation Transduction, Genetic Genetics Germ Cell Growth, Development And Aging Larva Male Metabolism Micro-injection Mutation Embryo, Nonmammalian Procedures Transgenic Animals Transplantation Vascularization Zygote Adult Germline Stem Cells Animal Animal, Genetically Modified Cichlids Embryo, Nonmammalian Female Gene Expression Regulation, Developmental Germ Cells Larva Male Microinjections Mutation Neovascularization, Physiologic Phosphatidylinositol 3-kinases Transduction, Genetic Zygote |
description |
Microinjection is commonly performed to achieve fish transgenesis; however, due to difficulties associated with this technique, new strategies are being developed. Here we evaluate the potential of lentiviral particles to genetically modify Nile tilapia cells to achieve transgenesis using three different approaches: spermatogonial stem cell (SSC) genetic modification and transplantation (SC), in vivo transduction of gametes (GT), and fertilised egg transduction (ET). The SC protocol using larvae generates animals with sustained production of modified sperm (80% of animals with 77% maximum sperm fluorescence [MSF]), but is a time-consuming protocol (sexual maturity in Nile tilapia is achieved at 6 months of age). GT is a faster technique, but the modified gamete production is temporary (70% of animals with 52% MSF). ET is an easier way to obtain mosaic transgenic animals compared to microinjection of eggs, but non-site-directed integration in the fish genome can be a problem. In this study, PI3Kc2α gene disruption impaired development during the embryo stage and caused premature death. The manipulator should choose a technique based on the time available for transgenic obtainment and if this generation is required to be continuous or not. © The Author(s) 2017. |
publishDate |
2017 |
dc.date.issued.fl_str_mv |
2017 |
dc.date.accessioned.fl_str_mv |
2020-05-07T14:14:47Z |
dc.date.available.fl_str_mv |
2020-05-07T14:14:47Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://repositorio.inpa.gov.br/handle/1/15219 |
dc.identifier.doi.none.fl_str_mv |
10.1038/srep44317 |
url |
https://repositorio.inpa.gov.br/handle/1/15219 |
identifier_str_mv |
10.1038/srep44317 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.pt_BR.fl_str_mv |
Volume 7 |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nc-nd/3.0/br/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nc-nd/3.0/br/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Scientific Reports |
publisher.none.fl_str_mv |
Scientific Reports |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional do INPA instname:Instituto Nacional de Pesquisas da Amazônia (INPA) instacron:INPA |
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INPA |
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Repositório Institucional do INPA |
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Repositório Institucional do INPA |
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