Department of Chemistry and Konstanz Research School Chemical Biology, University of Konstanz, Konstanz, Germany.
Methods Mol Biol. 2021;2323:199-212. doi: 10.1007/978-1-0716-1499-0_14.
Artificial RNA switches are an emerging class of genetic controllers suitable for synthetic biology applications. Aptazymes are fusions composed of an aptamer domain and a self-cleaving ribozyme. The utilization of aptazymes for conditional gene expression displays several advantages over employing conventional transcription factor-based techniques as aptazymes require minimal genomic space, fulfill their function without the need of protein cofactors and most importantly are reprogrammable with respect to ligand selectivity and the RNA function to be regulated. Technologies that enable the generation of aptazymes to defined input ligands are of interest for the construction of biocomputing devices and biosensing applications. In this chapter we present a method that facilitates the in vivo screening of randomized pools of aptazymes in mammalian cells.
人工 RNA 开关是一类新兴的遗传控制器，适用于合成生物学应用。适体酶是由适体结构域和自我切割核酶组成的融合蛋白。与传统的基于转录因子的技术相比，适体酶在条件性基因表达中的应用具有几个优势，因为适体酶需要最小的基因组空间，无需蛋白质辅助因子即可发挥作用，最重要的是，它们可以针对配体选择性和要调节的 RNA 功能进行重新编程。能够产生针对特定输入配体的适体酶的技术对于构建生物计算设备和生物传感应用具有重要意义。在本章中，我们介绍了一种在哺乳动物细胞中进行随机适体酶库体内筛选的方法。
