An artificial promoter construct for heat-inducible misexpression during fish embryogenesis

Developmental Biology
Baubak BajoghliThomas Czerny

Abstract

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...Continue Reading

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Citations

Oct 13, 2006·Molecular Biology of the Cell·Matthias Nowak, Matthias Hammerschmidt
Oct 25, 2006·Molecular and Cellular Biology·Thomas HeimbucherThomas Czerny
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