PUBLICATION
Transgenesis in fish: efficient selection of transgenic fish by co-injection with a fluorescent reporter construct
- Authors
- Rembold, M., Lahiri, K., Foulkes, N.S., and Wittbrodt, J.
- ID
- ZDB-PUB-070409-18
- Date
- 2006
- Source
- Nature Protocols 1(3): 1133-1139 (Journal)
- Registered Authors
- Foulkes, Nicholas-Simon, Lahiri, Kajori, Rembold, Martina, Wittbrodt, Jochen
- Keywords
- none
- MeSH Terms
-
- Animals
- Animals, Genetically Modified/genetics*
- Deoxyribonucleases, Type II Site-Specific
- Gene Expression*
- Gene Transfer Techniques*
- Green Fluorescent Proteins/metabolism
- Oryzias/genetics*
- Plasmids/genetics
- Saccharomyces cerevisiae Proteins
- Transgenes/genetics*
- Zebrafish/genetics*
- PubMed
- 17406394 Full text @ Nat. Protoc.
Citation
Rembold, M., Lahiri, K., Foulkes, N.S., and Wittbrodt, J. (2006) Transgenesis in fish: efficient selection of transgenic fish by co-injection with a fluorescent reporter construct. Nature Protocols. 1(3):1133-1139.
Abstract
Small fish are a popular laboratory model for studying gene expression and function by transgenesis. If, however, the transgenes are not readily detectable by visual inspection, a large number of embryos must be injected, raised and screened to identify positive founder fish. Here, we describe a strategy to efficiently generate and preselect transgenic lines harbouring any transgene of interest. Co-injection of a selectable reporter construct (e.g., GFP), together with the transgene of interest on a separate plasmid using the I-SceI meganuclease approach, results in co-distribution of the two plasmids. The quality of GFP expression within the F0 generation therefore reflects the quality of injection and allows efficient and reliable selection of founder fish that are also positive for the second transgene of interest. In our experience, a large fraction (up to 50%) of GFP-positive fish will also be transgenic for the second transgene, thus providing a rapid (within 3-4 months) and efficient way to establish transgenic lines for any gene of interest in medaka and zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping