Churgin Matthew A, He Liping, Murray John I, Fang-Yen Christopher
Department of Bioengineering, School of Engineering and Applied Sciences, University of Pennsylvania, Philadelphia, Pennsylvania 19104.
G3 (Bethesda). 2013 Oct 3;3(10):1827-32. doi: 10.1534/g3.113.007682.
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.
将转基因与热休克启动子偶联是一种广泛应用于调控基因表达的方法。在秀丽隐杆线虫中，基因诱导可通过热休克的时间控制来实现，也可通过在热休克突变体中进行靶向拯救来实现空间控制。在此，我们展示了一种能够以单细胞分辨率在任意细胞中引发基因表达的方法。我们使用聚焦脉冲红外激光在特定细胞中局部诱导热休克反应。我们的方法基于并扩展了先前报道的使用连续波激光的方法。在我们的技术中，由于脉冲之间的热扩散，脉冲激光照射能够实现更高程度的空间选择性。我们应用该方法在胚胎、幼虫和成虫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，是模型生物生物学的有力工具。
