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Targeted gene expression without a tissue-specific promoter: Creating mosaic embryos using laser-induced single-cell heat shock

By Marc S. Halfon, Hiroyuki Kose, Akira Chiba and Haig Keshishian

Abstract

We have developed a method to target gene expression in the Drosophila embryo to a specific cell without having a promoter that directs expression in that particular cell. Using a digitally enhanced imaging system to identify single cells within the living embryo, we apply a heat shock to each cell individually by using a laser microbeam. A 1- to 2-min laser treatment is sufficient to induce a heat-shock response but is not lethal to the heat-shocked cells. Induction of heat shock was measured in a variety of cell types, including neurons and somatic muscles, by the expression of β-galactosidase from an hsp26-lacZ reporter construct or by expression of a UAS target gene after induction of hsGAL4. We discuss the applicability of this technique to ectopic gene expression studies, lineage tracing, gene inactivation studies, and studies of cells in vitro. Laser heat shock is a versatile technique that can be adapted for use in a variety of research organisms and is useful for any studies in which it is desirable to express a given gene in only a distinct cell or clone of cells, either transiently or constitutively, at a time point of choice

Topics: Biological Sciences
Publisher: The National Academy of Sciences of the USA
Year: 1997
DOI identifier: 10.1073/pnas.94.12.6255
OAI identifier: oai:pubmedcentral.nih.gov:21036
Provided by: PubMed Central
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