Proteo-Science Center, Ehime University, 3 Bunkyo, Matsuyama, Ehime 790-8577, Japan.
Proteo-Science Center, Ehime University, 3 Bunkyo, Matsuyama, Ehime 790-8577, Japan.
Bioorg Med Chem Lett. 2022 Sep 1;71:128839. doi: 10.1016/j.bmcl.2022.128839. Epub 2022 May 30.
We chose two types of mid-sized Arg-rich peptides (Rev-pep and Tat-pep) as ligands and used their aptamers to construct efficient eukaryotic ON-riboswitches (ligand-dependently upregulating riboswitches). Due to the aptamers' high affinities, the best Rev-pep-responsive and Tat-pep-responsive riboswitches obtained showed much higher switching efficiencies at low ligand concentrations than small ligand-responsive ON-riboswitches in the same mechanism. In addition, despite the high sequence similarity of Rev-pep and Tat-pep, the two best riboswitches were almost insensitive to each other's peptide ligand. Considering the high responsiveness and specificity along with the versatility of the expression platform used and the applicability of Arg-rich peptides, this orthogonal pair of riboswitches would be widely useful eukaryotic gene regulators or biosensors.
我们选择了两种类型的中型富含精氨酸的肽（Rev-pep 和 Tat-pep）作为配体，并使用它们的适体构建高效的真核 ON-riboswitches（配体依赖性上调 riboswitches）。由于适体具有高亲和力，因此在低浓度配体下，获得的最佳 Rev-pep 响应和 Tat-pep 响应 riboswitches 的切换效率比相同机制中的小分子配体响应的 ON-riboswitches 高得多。此外，尽管 Rev-pep 和 Tat-pep 具有很高的序列相似性，但这两种最佳 riboswitches 对彼此的肽配体几乎不敏感。考虑到所使用的表达平台的高响应性和特异性以及通用性以及富含精氨酸的肽的适用性，这对正交 riboswitches 将成为广泛有用的真核基因调节剂或生物传感器。
