Lynch Sean A, Gallivan Justin P
Department of Chemistry, Emory University, Atlanta, GA 30322, USA.
Nucleic Acids Res. 2009 Jan;37(1):184-92. doi: 10.1093/nar/gkn924. Epub 2008 Nov 25.
Riboswitches regulate gene expression through direct, small molecule-mRNA interactions. The creation of new synthetic riboswitches from in vitro selected aptamers benefits from rapid, high-throughput methods for identifying switches capable of triggering dramatic changes in gene expression in the presence of a desired ligand. Here we present a flow cytometry-based screen for identifying synthetic riboswitches that induce robust increases in gene expression in the presence of theophylline. The performance characteristics of our newly identified riboswitches exceed those of previously described natural and synthetic riboswitches. Sequencing data and structure probing experiments reveal the ribosome binding site to be an important determinant of how well a switch performs and may provide insights into the design of new synthetic riboswitches.
核糖开关通过直接的小分子与mRNA相互作用来调节基因表达。从体外筛选的适配体创建新的合成核糖开关，受益于快速、高通量的方法，用于识别在所需配体存在下能够触发基因表达显著变化的开关。在这里，我们展示了一种基于流式细胞术的筛选方法，用于识别在茶碱存在下能诱导基因表达强劲增加的合成核糖开关。我们新鉴定的核糖开关的性能特征超过了先前描述的天然和合成核糖开关。测序数据和结构探测实验表明，核糖体结合位点是开关性能好坏的重要决定因素，可能为新合成核糖开关的设计提供见解。
