利用核糖开关的环状排列设计荧光生物传感器。
Designing fluorescent biosensors using circular permutations of riboswitches.
机构信息
Department of Chemistry, University of California, Berkeley 94720, USA.
College of Chemical and Molecular Engineering, Peking University, Beijing 100871, China.
出版信息
Methods. 2018 Jul 1;143:102-109. doi: 10.1016/j.ymeth.2018.02.014. Epub 2018 Feb 16.
Abstract
RNA-based fluorescent (RBF) biosensors have been applied to detect a variety of metabolites in vitro and in live cells. They are designed by combining the ligand sensing domain of natural riboswitches with in vitro selected fluorogenic aptamers. Different biosensor topologies have been developed to accommodate the diversity of riboswitch structures. Here we show that circular permutation of the riboswitch ligand sensing domain also gives functional biosensors, using the SAM-I riboswitch as our model. We reveal that this design can enhance fluorescence turn-on and ligand binding affinity compared to the non-permuted topology.
摘要
基于 RNA 的荧光（RBF）生物传感器已被应用于检测体外和活细胞中的各种代谢物。它们是通过将天然核酶的配体感应结构域与体外选择的荧光适体结合而设计的。为了适应核酶结构的多样性，已经开发了不同的生物传感器拓扑结构。在这里，我们展示了通过对核酶配体感应结构域进行环化排列，也可以得到功能性的生物传感器，我们选择 SAM-I 核酶作为模型。我们揭示了与非环化结构相比，这种设计可以增强荧光开启和配体结合亲和力。
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