Transactivation from Gal4-VP16 transgenic insertions for tissue-specific cell labeling and ablation in zebrafish
Transactivation from Gal4-VP16 transgenic insertions for tissue-specific cell labeling and ablation in zebrafish
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DOI：
10.1016/j.ydbio.2007.01.033
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发表时间：
2007-04-15
影响因子：
2.7
通讯作者：
Parsons, Michael J.
中科院分区：
文献类型：
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作者：
Davison, Jon M.;Akitake, Courtney M.;Parsons, Michael J.
先前对转基因斑马鱼的研究证实了转录因子Ga14在调节上游激活因子序列（UAS）下驱动其他基因表达的功能。然而，这种强大的二元系统的广泛应用部分受到相对低效的技术来建立转基因斑马鱼的效率低下，并且通过GA14的不足来影响UAS调节基因的高水平表达。我们已经使用TOL2换位系统在整个斑马鱼基因组中有效地分布自我报告的基因/增强子陷阱载体。该矢量使用有效的杂种转录因子GAL4-VP16激活UAS的表达：EGFP记者盒。在试点屏幕中，建立了稳定的转基因线，以可再现的模式表达EGFP，其中包括各种组织，包括大脑，脊髓，视网膜，诺科德，颅骨骨架和肌肉，并可以反式激活其他UAS受调查的基因。我们证明了这种方法在UAS中跟踪表达迁移细胞的GAL4-VP16的实用性。-KAEDE转基因鱼类，并使用UAS对照的细菌硝化还原酶基因诱导组织特异性细胞死亡。 TOL2介导的基因/增强子捕获系统与UAS转基因系一起为调节基因表达以及在斑马鱼早期发育期间特定细胞类型和组织的靶向标记和消融提供了有价值的工具。 （c）2007 Elsevier Inc.保留所有权利。
Prior studies with transgenic zebrafish confirmed the functionality of the transcription factor Ga14 to drive expression of other genes under the regulation of upstream activator sequences (UAS). However, widespread application of this powerful binary system has been limited, in part, by relatively inefficient techniques for establishing transgenic zebrafish and by the inadequacy of Ga14 to effect high levels of expression from UAS-regulated genes. We have used the Tol2 transposition system to distribute a self-reporting gene/enhancer trap vector efficiently throughout the zebrafish genome. The vector uses the potent, hybrid transcription factor Gal4-VP16 to activate expression from a UAS:eGFP reporter cassette. In a pilot screen, stable transgenic lines were established that express eGFP in reproducible patterns encompassing a wide variety of tissues, including the brain, spinal cord, retina, notochord, cranial skeleton and muscle, and can transactivate other UAS-regulated genes. We demonstrate the utility of this approach to track Gal4-VP16 expressing migratory cells in UAS.-Kaede transgenic fish, and to induce tissue-specific cell death using a bacterial nitroreductase gene under UAS control. The Tol2-mediated gene/enhancer trapping system together with UAS transgenic lines provides valuable tools for regulated gene expression and for targeted labeling and ablation of specific cell types and tissues during early zebrafish development. (c) 2007 Elsevier Inc. All rights reserved.