Alteration of Drosophila life span using conditional, tissue-specific expression of transgenes triggered by doxycyline or RU486/Mifepristone
Alteration of Drosophila life span using conditional, tissue-specific expression of transgenes triggered by doxycyline or RU486/Mifepristone
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DOI：
10.1016/j.exger.2007.01.004
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发表时间：
2007-06-01
影响因子：
3.9
通讯作者：
Tower, John
中科院分区：
文献类型：
--
作者：
Ford, Daniel;Hoe, Nicholas;Tower, John
比较和优化了Tet-on和Geneswitch两种条件系统，用于转基因的组织特异性表达和寿命调控。比较了Tet-on系统反向四环素反式激活因子（rtTA）的两个版本：原始rtTA和包含旨在优化调控和表达的突变的rtTAM 2-alt。rtTAM 2-alt版本在不存在多西环素的情况下产生较少的靶构建体的泄漏表达，然而，与先前的报道相反，可以实现的表达的绝对水平低于rtTA产生的表达水平。将现有的UAS-rtTAM 2-alt插入重新平衡，并与几种组织通用和组织特异性GAL 4驱动系组合，以产生超过三个数量级的组织特异性、多西环素诱导的转基因表达。Geneswitch（GS）系统也具有低背景，但相对于Tet-on系统，绝对表达水平较低。因此，产生了actin 5C-GS多插入染色体，并在不增加背景的情况下实现了更高水平的表达。MnSOD的适度过表达对寿命有有益影响。在这里，MnSOD的高水平过表达被发现具有毒性作用。相反，运动神经元特异性MnSOD的过度表达对寿命没有可检测的影响。结果表明，运动神经元组织不是MnSOD诱导的寿命或毒性的成年男性的基本组织。(c)2007年爱思唯尔公司All rights reserved.
The conditional systems Tet-on and Geneswitch were compared and optimized for the tissue-specific expression of transgenes and manipulation of life span in adult Drosophila. Two versions of Tet-on system reverse-tetracycline-Trans-Activator (rtTA) were compared: the original rtTA, and rtTAM2-alt containing mutations designed to optimize regulation and expression. The rtTAM2-alt version gave less leaky expression of target constructs in the absence of doxycyline, however the absolute level of expression that could be achieved was less than that produced by rtTA, in contrast to a previous report. Existing UAS-rtTAM2-alt insertions were re-balanced, and combined with several tissue-general and tissue-specific GAL4 driver lines to yield tissue-specific, doxycyline-inducible transgene expression over three orders of magnitude. The Geneswitch (GS) system also had low background, but the absolute level of expression was low relative to Tet-on. Consequently, actin5C-GS multi-insert chromosomes were generated and higher-level expression was achieved without increased background. Moderate level over-expression of MnSOD has beneficial effects on life span. Here high-level over-expression of MnSOD was found to have toxic effects. In contrast, motor-neuron-specific over-expression of MnSOD had no detectable effect on life span. The results suggest that motor-neuron tissue is not the essential tissue for either MnSOD induced longevity or toxicity in adult males. (c) 2007 Elsevier Inc. All rights reserved.