Spatiotemporal gene expression targeting with the TARGET and gene-switch systems in Drosophila.
Spatiotemporal gene expression targeting with the TARGET and gene-switch systems in Drosophila.
复制标题
DOI：
10.1126/stke.2202004pl6
复制
发表时间：
2004-02-12
期刊：
影响因子：
--
通讯作者：
Davis, Ronald L
中科院分区：
文献类型：
--
作者：
McGuire, Sean E;Mao, Zhengmei;Davis, Ronald L
靶向基因表达已成为研究果蝇生物学问题的一种标准技术。直到最近，转基因表达只能在时间或空间维度上进行靶向，而不能同时在两个维度上进行。最近已经开发了几种新的系统，可以同时在时间和空间上引导转基因表达。我们在此描述我们实验室开发的两种这样的系统。第一种系统利用常规的GAL4 - 上游激活序列（UAS）系统和一种温度敏感的GAL80分子，提供了一种针对时间和区域的基因表达靶向（TARGET）的通用方法，该GAL80分子在允许的温度下抑制GAL4的转录活性。第二种系统称为基因开关（Gene - Switch），基于一种可被激素诱导的GAL4 - 孕酮受体嵌合体。我们已经使用这两种系统同时在空间和时间上挽救果蝇芜菁（rutabaga，rut）记忆突变体的记忆功能障碍。在本实验方案中，我们提供了使用这两种新系统的指南，这两种系统在研究果蝇生物学以及将果蝇用作人类疾病模型方面应该具有普遍的实用性。
Targeted gene expression has become a standard technique for the study of biological questions in Drosophila. Until recently, transgene expression could be targeted in the dimension of either time or space, but not both. Several new systems have recently been developed to direct transgene expression simultaneously in both time and space. We describe here two such systems that we developed in our laboratory. The first system provides a general method for temporal and regional gene expression targeting (TARGET) with the conventional GAL4-upstream activator sequence (UAS) system and a temperature-sensitive GAL80 molecule, which represses GAL4 transcriptional activity at permissive temperatures. The second system, termed Gene-Switch, is based on a GAL4-progesterone receptor chimera that is hormone-inducible. We have used both systems for simultaneous spatial and temporal rescue of memory dysfunction in the rutabaga (rut) memory mutant of Drosophila. In this protocol, we provide guidelines for the use of these two novel systems, which should have general utility in studying Drosophila biology and in using the fly as a model for human disease.