Hardy Melissa E, Ross Louis V, Chien Chi-Bin
Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, Utah, USA.
Dev Dyn. 2007 Nov;236(11):3071-6. doi: 10.1002/dvdy.21318.
Misexpression of genes in a temporally and spatially controlled fashion is an important tool for assessing gene function during development. Because few tissue-specific promoters have been identified in zebrafish, inducible systems such as the Cre/LoxP and Tet repressor systems are of limited utility. Here we describe a new method of misexpression: local heat shock using a modified soldering iron. Zebrafish carrying transgenes under the control of a heat shock promoter (hsp70) are focally heated with the soldering iron to induce gene expression in a small area of the embryo. We have validated this method in three stable transgenic lines and at three developmental timepoints. Local heat shock is a fast, easy, and inexpensive method for gene misexpression.
以时间和空间可控的方式错误表达基因是评估发育过程中基因功能的重要工具。由于在斑马鱼中鉴定出的组织特异性启动子很少，Cre/LoxP和Tet阻遏系统等诱导系统的实用性有限。在此，我们描述一种新的错误表达方法：使用改良烙铁进行局部热休克。携带在热休克启动子（hsp70）控制下的转基因的斑马鱼用烙铁进行局部加热，以在胚胎的小区域诱导基因表达。我们已在三个稳定转基因系和三个发育时间点验证了该方法。局部热休克是一种用于基因错误表达的快速、简便且廉价的方法。
