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Gene delivery to Nile tilapia cells for transgenesis and the role of PI3K-c2α in angiogenesis
- Source :
- Scientific Reports, Repositório Institucional do INPA, Instituto Nacional de Pesquisas da Amazônia (INPA), instacron:INPA
- Publication Year :
- 2017
- Publisher :
- Springer Science and Business Media LLC, 2017.
-
Abstract
- 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.
- Subjects :
- Male
0301 basic medicine
Embryo, Nonmammalian
Animal, Genetically Modified
Zygote
Phosphatidylinositol 3 Kinase
Procedures
Animals, Genetically Modified
Phosphatidylinositol 3-Kinases
Nile tilapia
0302 clinical medicine
Transduction, Genetic
Multidisciplinary
Adult Germline Stem Cells
biology
Gene Expression Regulation, Developmental
Cichlids
Micro-injection
Cell biology
Transgenesis
medicine.anatomical_structure
Larva
Cichlid
Deficiency
Gamete
Female
Transgenic Animals
Phosphatidylinositol 3-kinases
Microinjections
Transgene
Neovascularization, Physiologic
Gene delivery
Germ Cell
Article
03 medical and health sciences
Genetics
medicine
Animals
Microinjection
Transplantation
Animal
Vascularization
biology.organism_classification
Sperm
Molecular biology
Metabolism
Germ Cells
030104 developmental biology
Gene Expression Regulation
Growth, Development And Aging
Adult Stem Cell
Mutation
Angiogenesis
Cytology
030217 neurology & neurosurgery
Subjects
Details
- ISSN :
- 20452322
- Volume :
- 7
- Database :
- OpenAIRE
- Journal :
- Scientific Reports
- Accession number :
- edsair.doi.dedup.....7e7ad648eb912b48d3b95795463f7d0a