An artificial promoter construct for heat-inducible misexpression during fish embryogenesis
- Authors
- Bajoghli, B., Aghaallaei, N., Heimbucher, T., and Czerny, T.
- ID
- ZDB-PUB-110701-11
- Date
- 2004
- Source
- Developmental Biology 271(2): 416-430 (Journal)
- Registered Authors
- Aghaallaei, Narges, Bajoghli, Baubak
- Keywords
- heat shock promoter, inducible misexpression, medaka, injection, meganuclease, fgf8, cyclopic eye, otic vesicle
- MeSH Terms
-
- Mice
- Humans
- Transcriptional Activation
- Luminescent Proteins
- Heat-Shock Proteins/genetics*
- Time Factors
- Cloning, Molecular/methods*
- Fibroblast Growth Factor 8
- Animals
- Animals, Genetically Modified
- Green Fluorescent Proteins
- Fibroblast Growth Factors/genetics*
- Fibroblast Growth Factors/metabolism
- Oryzias/embryology*
- Oryzias/genetics
- Tumor Cells, Cultured
- Microinjections
- Hot Temperature*
- Promoter Regions, Genetic/genetics*
- Gene Expression Regulation, Developmental*
- Luciferases
- PubMed
- 15223344 Full text @ Dev. Biol.
Beside spatial distribution, timing of gene expression is a key parameter controlling gene function during embryonic development. Gain-of-function experiments can therefore have quite opposing results, depending on the time of gene activation. Induction techniques are necessary to control timing in these experiments from outside of the organism. Natural heat shock promoters constitute a simple inducible misexpression system, the main disadvantage is a high background level of expression. We present here a new heat stress-inducible bidirectional promoter consisting of multimerized heat shock elements (HSE). The simplified architecture of this promoter largely improves the properties needed for an efficient induction system: dramatically reduced background activity, improved inducibility, and loss of all tissue specific components. Based on this new artificial promoter, we present a transient induction system for fish embryos. Application of this new induction system for Fgf8 misexpression during embryonic development reveals different windows of competence during eye development. A dramatic early phenotype resulting in loss of the eyes is observed for conventional mRNA injection. Later activation, by using our inducible promoter, uncovers different eye phenotypes like cyclopic eyes. Even after 14 days, an efficient heat stress response could be evoked in the injected embryos. The HSE promoter therefore represents a new artificial heat shock promoter with superior properties, making possible transient experiments with inducible misexpression at various stages of development.