A high-throughput screen for synthetic riboswitches reveals mechanistic insights into their function
A high-throughput screen for synthetic riboswitches reveals mechanistic insights into their function
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DOI：
10.1016/j.chembiol.2006.12.008
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发表时间：
2007-02-01
影响因子：
--
通讯作者：
Gallivan, Justin P.
中科院分区：
文献类型：
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作者：
Lynch, Sean A.;Desai, Shawn K.;Gallivan, Justin P.
核糖开关是基于RNA的遗传控制元件，其以配体依赖的方式调节基因表达而不需要蛋白质。产生响应于任何所需的小分子配体控制基因表达的合成核糖开关的能力将使得能够开发可以检测小分子的存在的敏感的遗传筛选，以及设计遗传控制元件以有条件地调节细胞行为。在此，我们提出了一种自动化的高通量筛选方法，该方法鉴定了在不存在所需配体的情况下显示极低背景水平的基因表达和在其存在下表达的稳健增加的合成核糖开关。机制研究揭示了这些核糖开关的功能，并提出了创造新的合成核糖开关的设计原则。我们预计，筛选方法和设计原则将普遍适用于创建功能性合成核糖开关。
Riboswitches are RNA-based genetic control elements that regulate gene expression in a ligand-dependent fashion without the need for proteins. The ability to create synthetic riboswitches that control gene expression in response to any desired small-molecule ligand will enable the development of sensitive genetic screens that can detect the presence of small molecules, as well as designer genetic control elements to conditionally modulate cellular behavior. Herein, we present an automated high-throughput screening method that identifies synthetic riboswitches that display extremely low background levels of gene expression in the absence of the desired ligand and robust increases in expression in its presence. Mechanistic studies reveal how these riboswitches function and suggest design principles for creating new synthetic riboswitches. We anticipate that the screening method and design principles will be generally useful for creating functional synthetic riboswitches.