Waldner Christoph, Sakamaki Kazuhiro, Ueno Naoto, Turan Gülüzar, Ryffel Gerhart U
Institut für Zellbiologie (Tumorforschung), Universitätsklinikum Essen, Essen, Germany.
Dev Dyn. 2006 Aug;235(8):2220-8. doi: 10.1002/dvdy.20880.
For reproducible analyses of gene function in Xenopus, the use of transgenic strains is a promising approach but has limitations when investigating factors interfering with development. Therefore, inducible systems are attractive alternatives, and a binary system based on recombinases is a most versatile approach. We have shown previously that Cre and FLP recombinases are active in Xenopus laevis and can induce a silent reporter gene in a corresponding reporter strain. Here, we describe the establishment of the transgenic Xenopus laevis strain A7 expressing Cre recombinase under the control of the muscle-specific cardiac actin promoter. Upon crossing to several distinct reporter strains, A7 is able to induce EYFP, DsRed2, or LacZ reporter genes in a muscle-specific manner. This first Cre-expressing strain allows conditional activation of any gene of interest in muscle cells and, thus, opens up the use of recombinases as a new experimental strategy in Xenopus.
为了在非洲爪蟾中对基因功能进行可重复分析,使用转基因品系是一种很有前景的方法,但在研究干扰发育的因素时存在局限性。因此,诱导系统是有吸引力的替代方法,基于重组酶的二元系统是一种用途最为广泛的方法。我们之前已经表明,Cre和FLP重组酶在非洲爪蟾中具有活性,并且可以在相应的报告品系中诱导沉默的报告基因。在这里,我们描述了转基因非洲爪蟾品系A7的建立,该品系在肌肉特异性心肌肌动蛋白启动子的控制下表达Cre重组酶。与几个不同的报告品系杂交后,A7能够以肌肉特异性方式诱导EYFP、DsRed2或LacZ报告基因。这个首个表达Cre的品系允许在肌肉细胞中有条件地激活任何感兴趣的基因,从而开启了将重组酶作为非洲爪蟾新实验策略的应用。