PUBLICATION
Viral 2A peptides allow expression of multiple proteins from a single ORF in transgenic zebrafish embryos
- Authors
- Provost, E., Rhee, J., and Leach, S.D.
- ID
- ZDB-PUB-071023-8
- Date
- 2007
- Source
- Genesis (New York, N.Y. : 2000) 45(10): 625-629 (Journal)
- Registered Authors
- Leach, Steven D.
- Keywords
- 2A self-cleaving peptide, zebrafish, transgenesis, multicistronic, IRES
- MeSH Terms
-
- Embryo, Nonmammalian
- Genes, Reporter
- Transgenes
- Zebrafish Proteins/genetics*
- Zebrafish Proteins/metabolism
- Zebrafish/embryology
- Zebrafish/genetics*
- Peptides/genetics*
- Gene Expression Regulation, Developmental*
- DNA, Complementary
- Animals
- Animals, Genetically Modified
- Luminescent Proteins/metabolism
- Green Fluorescent Proteins/metabolism
- Open Reading Frames*
- PubMed
- 17941043 Full text @ Genesis
Citation
Provost, E., Rhee, J., and Leach, S.D. (2007) Viral 2A peptides allow expression of multiple proteins from a single ORF in transgenic zebrafish embryos. Genesis (New York, N.Y. : 2000). 45(10):625-629.
Abstract
We have adapted a novel multicistronic gene expression system involving viral peptides to the zebrafish. The viral 2A peptide allows production of multiple protein products from a single transgene. Based on highly inefficient peptide bond formation between glycine and proline residues within the 2A peptide, placement of 2A peptide sequence as a linker region between tandem cDNA's allows the stoichiometric translation of multiple unfused protein products. To test this system in zebrafish, we generated two different tandem reporter constructs employing eGFP and mCherry, separated by 2A peptide sequence. Using this system, we produced transgenic zebrafish in which fluorophores were produced as independent proteins from a single transcript. The successful application of this technology in zebrafish will be valuable for visually marking transgenic embryos and transgene-expressing cells, or in any situation where reliable expression of multiple transgenes is desired.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping