title: Efficient single-cell transgene induction in Caenorhabditis elegans using a pulsed infrared laser.
abstract: The coupling of transgenes to heat shock promoters is a widely applied method for regulating gene expression. In C. elegans, gene induction can be controlled temporally through timing of heat shock and spatially via targeted rescue in heat shock mutants. Here, we present a method for evoking gene expression in arbitrary cells, with single-cell resolution. We use a focused pulsed infrared laser to locally induce a heat shock response in specific cells. Our method builds on and extends a previously reported method using a continuous-wave laser. In our technique, the pulsed laser illumination enables a much higher degree of spatial selectivity because of diffusion of heat between pulses. We apply our method to induce transient and long-term transgene expression in embryonic, larval, and adult cells. Our method allows highly selective spatiotemporal control of transgene expression and is a powerful tool for model organism biology.
authors: Department of Bioengineering, School of Engineering and Applied Sciences, University of Pennsylvania, Philadelphia, Pennsylvania 19104.
authors: Matthew A
authors: MA
authors: Churgin
authors: Liping
authors: L
authors: He
authors: John I
authors: JI
authors: Murray
authors: Christopher
authors: C
authors: Fang-Yen
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics* / Q000378:metabolism; D017173:Caenorhabditis elegans / Q000235:genetics* / Q000378:metabolism; D018869:Heat-Shock Response; D006361:Heating / Q000379:methods; D007834:Lasers; D059010:Single-Cell Analysis / Q000379:methods; D014158:Transcription, Genetic
keywords: C. elegans; heat shock; transgene induction
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