PUBLICATION
Practical procedures for ectopic induction of gene expression in zebrafish embryos using Bhc-diazo-caged mRNA
- Authors
- Ando, H. and Okamoto, H.
- ID
- ZDB-PUB-031229-5
- Date
- 2003
- Source
- Methods in cell science : an official journal of the Society for In Vitro Biology 25(1-2): 25-31 (Review)
- Registered Authors
- Ando, Hideki, Okamoto, Hitoshi
- Keywords
- Bhc-diazo, Caged mRNA, Caging, Ectopic gene expression, Zebrafish
- MeSH Terms
-
- Animals
- Azo Compounds/chemistry*
- Cloning, Molecular
- Coumarins/chemistry*
- Embryo, Nonmammalian/anatomy & histology
- Microinjections
- Photolysis/radiation effects*
- RNA, Messenger/genetics
- RNA, Messenger/metabolism*
- Ultraviolet Rays
- Zebrafish/anatomy & histology
- Zebrafish/genetics*
- beta-Galactosidase/genetics
- beta-Galactosidase/metabolism*
- PubMed
- 14739584 Full text @ Methods Cell Sci.
Citation
Ando, H. and Okamoto, H. (2003) Practical procedures for ectopic induction of gene expression in zebrafish embryos using Bhc-diazo-caged mRNA. Methods in cell science : an official journal of the Society for In Vitro Biology. 25(1-2):25-31.
Abstract
We previously reported mRNA caging technology as a novel and simple technique for photo-mediated temporal and spatial control of gene activation in zebrafish embryos and as an alternative to the 'gene knockdown' approach using antisense morpholino oligonucleotides. The caging reagent used is 6-bromo-4-diazomethyl-7-hydroxycoumarin (Bhc-diazo), which forms a covalent bond with the phosphate moiety of the sugar-phosphate backbone of RNA. Mainly because of the reduced solubility of caged mRNA in aqueous solutions, special care in handling is needed. The Bhc-diazo group binds to the phosphate moieties of RNA and abolishes the translational activity of the latter. The translational activity of Bhc-caged mRNA is restored by photolysis/uncaging when exposed to long-wave UV light (350~365 nm). In this paper we describe the technique and detailed procedures for spatially and temporally controlled induction of gene expression in zebrafish embryos.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping