PUBLICATION
Improving the production of transgenic fish germlines: in vivo evaluation of mosaicism in zebrafish (Danio rerio) using a green fluorescent protein (GFP) and growth hormone cDNA transgene co-injection strategy
- Authors
- Figueiredo, M.A., Lanes, C.F.C., Almeida, D.V., and Marins, L.F.
- ID
- ZDB-PUB-100621-43
- Date
- 2007
- Source
- Genetics and Molecular Biology 30(1): 31-36 (Journal)
- Registered Authors
- Almeida, Daniela Volcan, Figueiredo, Marcio de Azevedo, Lanes, Carlos Frederico Ceccon, Marins, Luis Fernando
- Keywords
- transgenesis, genetically modified fish, microinjection, growth hormone cDNA
- MeSH Terms
- none
- PubMed
- none Full text @ Genet. Mol. Biol.
Citation
Figueiredo, M.A., Lanes, C.F.C., Almeida, D.V., and Marins, L.F. (2007) Improving the production of transgenic fish germlines: in vivo evaluation of mosaicism in zebrafish (Danio rerio) using a green fluorescent protein (GFP) and growth hormone cDNA transgene co-injection strategy. Genetics and Molecular Biology. 30(1):31-36.
Abstract
In fish, microinjection is the method most frequently used for gene transfer. However, due to delayed transgene integration this technique almost invariably produces mosaic individuals and if the gene is not integrated into germ cells its transmission to descendants is difficult or impossible. We evaluated the degree of in vivo mosaicism using a strategy where a reporter transgene is co-injected with a transgene of interest so that potential germline founders can be easily identified. Transgenic zebrafish (Danio rerio) were produced using two transgenes, both comprised of the carp b-actin promoter driving the expression of either the green fluorescent protein (GFP) reporter gene or the growth hormone cDNA from the marine silverside fish Odonthestes argentinensis. The methodology applied allowed a rapid identification of G0 transgenic fish and also detected which fish were transmitting transgenes to the next generation. This strategy also allowed inferences to be made about genomic transgene integration events in the six lineages produced and allowed the identification of one lineage transmitting both transgenes linked on the same chromosome. These results represent a significant advance in the reduction of the effort invested in producing a stable genetically modified fish lineage.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping