A modular and extensible RNA-based gene-regulatory platform for engineering cellular function
A modular and extensible RNA-based gene-regulatory platform for engineering cellular function
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DOI：
10.1073/pnas.0703961104
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发表时间：
2007-09-04
影响因子：
11.1
通讯作者：
Smolke, Christina D.
中科院分区：
文献类型：
--
作者：
Win, Maung Nyan;Smolke, Christina D.
工程生物系统有望解决人类在化学加工、能源生产、材料建设、维护和增强人类健康和环境方面的迫切需求。然而，我们设计生物系统的能力的重大进步受到了基础工具的限制，这些工具可用于报告、响应和控制生命系统中的细胞内成分。为了可靠地构建跨不同生物的通信和控制系统，需要便携式和可扩展的平台。我们报告了一个可扩展的基于rna的框架，用于工程配体控制的基因调控系统，称为核酶开关，它具有可调节的调节、设计模块化和目标特异性。这些开关平台包含一个传感器域，由适体序列组成，和一个执行器域，由锤头核酶序列组成。我们研究了这些结构域之间的两种标准化信息传递模式，并展示了一种机制，该机制允许可靠和模块化的功能合成RNA开关组装和对各种效应物的核酶活性调节。除了展示小分子反应，体内功能，变构锤头核酶的例子外，这项工作还描述了构建便携式和可扩展的基因调控系统的一般方法。我们展示了该平台在实现特定应用控制系统中的多功能性，用于小分子介导的细胞生长调节和无创体内代谢物产生的传感。
Engineered biological systems hold promise in addressing pressing human needs in chemical processing, energy production, materials construction, and maintenance and enhancement of human health and the environment. However, significant advancements in our ability to engineer biological systems have been limited by the foundational tools available for reporting on, responding to, and controlling intracellular components in living systems. Portable and scalable platforms are needed for the reliable construction of such communication and control systems across diverse organisms. We report an extensible RNA-based framework for engineering ligand-controlled gene-regulatory systems, called ribozyme switches, that exhibits tunable regulation, design modularity, and target specificity. These switch platforms contain a sensor domain, comprised of an aptamer sequence, and an actuator domain, comprised of a hammerhead ribozyme sequence. We examined two modes of standardized information transmission between these domains and demonstrate a mechanism that allows for the reliable and modular assembly of functioning synthetic RNA switches and regulation of ribozyme activity in response to various effectors. In addition to demonstrating examples of small molecule-responsive, in vivo functional, allosteric hammerhead ribozymes, this work describes a general approach for the construction of portable and scalable gene-regulatory systems. We demonstrate the versatility of the platform in implementing application-specific control systems for small molecule-mediated regulation of cell growth and noninvasive in vivo sensing of metabolite production.