Gene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesis
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 diff...
| Autor principal: | Tonelli, Fernanda MP |
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| Outros Autores: | dos Santos NassifLacerda, Samyra Maria, Procópio, Marcela Santos, Lemos, Breno Luiz Sales, França, Luiz Renato de, Resende, Rodrigo Ribeiro |
| Grau: | Artigo |
| Idioma: | English |
| Publicado em: |
Scientific Reports
2020
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| Acesso em linha: |
https://repositorio.inpa.gov.br/handle/1/15219 |
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oai:repositorio:1-15219 |
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oai:repositorio:1-15219 Gene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesis Tonelli, Fernanda MP dos Santos NassifLacerda, Samyra Maria Procópio, Marcela Santos Lemos, Breno Luiz Sales França, Luiz Renato de Resende, Rodrigo Ribeiro 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 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. 2020-05-07T14:14:47Z 2020-05-07T14:14:47Z 2017 Artigo https://repositorio.inpa.gov.br/handle/1/15219 10.1038/srep44317 en Volume 7 Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nc-nd/3.0/br/ application/pdf Scientific Reports |
| institution |
Instituto Nacional de Pesquisas da Amazônia - Repositório Institucional |
| collection |
INPA-RI |
| language |
English |
| 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 |
| spellingShingle |
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 Tonelli, Fernanda MP Gene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesis |
| topic_facet |
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. |
| format |
Artigo |
| author |
Tonelli, Fernanda MP |
| author2 |
dos Santos NassifLacerda, Samyra Maria Procópio, Marcela Santos Lemos, Breno Luiz Sales França, Luiz Renato de Resende, Rodrigo Ribeiro |
| author2Str |
dos Santos NassifLacerda, Samyra Maria Procópio, Marcela Santos Lemos, Breno Luiz Sales França, Luiz Renato de Resende, Rodrigo Ribeiro |
| title |
Gene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesis |
| 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 |
| publisher |
Scientific Reports |
| publishDate |
2020 |
| url |
https://repositorio.inpa.gov.br/handle/1/15219 |
| _version_ |
1787142083828514816 |
| score |
11.653393 |