PubAg

Main content area

Designing fluorescent biosensors using circular permutations of riboswitches

Author:
Truong, Johnny, Hsieh, Yu-Fang, Truong, Lynda, Jia, Guifang, Hammond, Ming C.
Source:
Methods 2018 v.143 pp. 102-109
ISSN:
1046-2023
Subject:
binding capacity, biosensors, fluorescence, ligands, metabolites, models, oligonucleotides, topology
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.
Agid:
6118207