Engineering of designer switches for controlling protein expression in eukaryotic model organisms (A05)

在第二个资助期内，我们成功地实现了基于RNA的基因表达开关，允许在模式生物C中诱导基因表达。优雅所开发的开关使得能够实现亨廷顿病模型，由于易于聚集的亨廷顿蛋白变体的细胞毒性，该模型在组成型设置中是不可能的。在第三个资助期内，我们将优化开关的动态范围，并将OFF开关添加到C中可用的曲目中。优雅此外，我们将探讨是否可以利用额外的配体在线虫。除了利用核酶作为表达平台外，我们还将开发其他在基因开关中实现RNA适体的方法，例如控制剪接位点或poly-A信号可及性。所开发的RNA开关将用于控制C. elegans与E. Deuerling和M. Gamerdinger（项目A01和A07），与M. Groettrup（项目C 01）和A. thaliana与E. Isono（项目C 08）。

Within the second funding period we have successfully implemented RNA-based switches of gene expression allowing for induction of gene expression in the model organism C. elegans. The developed switches enabled the implementation of a Huntington´s Disease model that would not have been possible in a constitutive setup due to the cytotoxicity of the aggregation-prone Huntingtin variants. Within the third funding period we will optimize the dynamic range of the switches and add OFF-switches to the repertoire available in C. elegans. Furthermore, we will explore whether additional ligands can be utilized in the nematode. In addition to utilizing ribozymes as expression platforms, we will develop other ways of implementing RNA aptamers in genetic switches such as controlling splice site or poly-A signal accessibility. The developed RNA switches will be applied for controlling gene expression in C. elegans in collaborative efforts with E. Deuerling and M. Gamerdinger (projects A01 and A07), in human cell lines together with M. Groettrup (project C01), and in A. thaliana with E. Isono (project C08).