論文 - 詳細
| RRC ID | 376 |
|---|---|
| 著者 | Ando H, Okamoto H. |
| タイトル | Practical procedures for ectopic induction of gene expression in zebrafish embryos using Bhc-diazo-caged mRNA. |
| ジャーナル | Methods Cell Sci |
| 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 approximately 365 nm). In this paper we describe the technique and detailed procedures for spatially and temporally controlled induction of gene expression in zebrafish embryos. |
| 巻・号 | 25(1-2) |
| ページ | 25-31 |
| 公開日 | 2003-1-1 |
| DOI | 10.1023/B:MICS.0000006846.13226.38 |
| PII | 5146565 |
| PMID | 14739584 |
| MeSH | 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* |
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