A novel Cre/loxP system for mosaic gene expression in the Drosophila embryo
A novel Cre/loxP system for mosaic gene expression in the Drosophila embryo
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DOI：
10.1002/dvdy.23784
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发表时间：
2012-05-01
影响因子：
2.5
通讯作者：
Matsuno, Kenji
中科院分区：
文献类型：
--
作者：
Nakazawa, Naotaka;Taniguchi, Kiichiro;Matsuno, Kenji
背景资料：镶嵌分析用于评估生物体中细胞亚群的基因功能和细胞自主性，并已广泛应用于果蝇研究。然而，利用现有的方法很难在果蝇胚胎组织中产生嵌合细胞。因此，我们开发了一种新的方法来产生遗传嵌合体胚胎使用修改的Cre/loxP系统。在本报告中，我们还描述了这种新方法的能力和局限性。结果如下：我们首先构建了一个新的盒，结合loxP与肌动蛋白5C增强子和Gal 4 cDNA，并产生了一个转基因苍蝇携带这个构建体（Aloxg-Gal 4）。在Aloxg-Gal 4中，Gal 4表达的激活被吉普赛绝缘子抑制。然而，一旦去除吉普赛绝缘子，当Cre重组酶诱导loxP位点之间的位点特异性重组时，Gal 4表达。该系统允许Gal 4在果蝇胚胎组织（表皮、浆膜、气管系统、马氏管、前肠、后肠、中肠和神经元）中嵌合表达，导致在上游激活序列（UAS）控制下的任意基因的Gal 4依赖性激活。结论：这种实用的方法可用于在果蝇胚胎组织中产生镶嵌细胞，并且可以应用于任何基因，而无需专门的设备。发展动力学241：965974，2012年。(c)2012 Wiley Periodicals，Inc.
Background: Mosaic analysis is used to assess gene function and cell autonomy in a subset of cells in an organism, and has been extensively applied in Drosophila studies. However, it is difficult to generate mosaic cells in Drosophila embryonic tissues using existing methods. Therefore, we developed a new method for generating genetic mosaic embryos using a modified Cre/loxP system. In this report, we also characterized the capabilities and limitations of this novel method. Results: We first constructed a novel cassette combining loxP with the Actin 5C enhancer and Gal4 cDNA, and generated a transgenic fly carrying this construct (Aloxg-Gal4). In Aloxg-Gal4, the activation of Gal4 expression is suppressed by the gypsy insulator. Once the gypsy insulator is removed, however, Gal4 is expressed when site-specific recombination between loxP sites is induced by Cre recombinase. This system allowed the mosaic expression of Gal4 in Drosophila embryonic tissues (epidermis, amnioserosa, tracheal system, malpighian tubules, foregut, hindgut, midgut, and neuron), leading to the Gal4-dependent activation of arbitrary genes under the control of the upstream activation sequence (UAS). Conclusions: This practical method can be used to generate mosaic cells in Drosophila embryonic tissues and can be applied to any gene without specialized equipment. Developmental Dynamics 241:965974, 2012. (c) 2012 Wiley Periodicals, Inc.