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

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

Research output: Contribution to journalArticle

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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 n 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 hsp26-1acZ 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.

Original languageEnglish (US)
Pages (from-to)6255-6260
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume94
Issue number12
DOIs
StatePublished - Jun 10 1997
Externally publishedYes

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