Efficient single-cell transgene induction in Caenorhabditis elegans using a pulsed infrared laser.
Efficient single-cell transgene induction in Caenorhabditis elegans using a pulsed infrared laser.
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DOI：
10.1534/g3.113.007682
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发表时间：
2013-10-03
期刊：
影响因子：
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通讯作者：
Fang-Yen C
中科院分区：
文献类型：
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作者：
Churgin MA;He L;Murray JI;Fang-Yen C
转基因与热休克启动子的偶联是一种广泛应用的基因表达调控方法。在线虫中，基因诱导可以通过热休克的时机在时间上进行控制，在热休克突变体中通过靶向抢救来进行空间控制。在这里，我们提出了一种在任意细胞中激发基因表达的方法，具有单细胞分辨率。我们使用聚焦的脉冲红外激光在特定细胞中局部诱导热休克反应。我们的方法建立在以前报道的使用连续波激光的方法的基础上，并进行了扩展。在我们的技术中，由于脉冲之间的热扩散，脉冲激光照明能够实现更高程度的空间选择性。我们应用我们的方法在胚胎、幼虫和成体细胞中诱导瞬时和长期转基因表达。我们的方法允许对转基因表达进行高度选择性的时空控制，是模式生物生物学的有力工具。
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.