Riboswitch-sRNA for Dual Transcript Control by a Ligand

用于通过配体控制双转录的核糖开关-sRNA

• 批准号: 1158394
• 负责人: Richard Lease
• 金额: $ 40万
• 依托单位: Ohio State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-15 至 2016-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1158394&HistoricalAwards=false
• 关键词: Riboswitch; sRNA; Dual; Transcript; Control

1158394/ LeaseThe capacity to reprogram bacteria to carry out useful tasks builds upon existing genetic circuit elements that can be repurposed. Two challenges to the design of synthetic bio-circuits include coordination of expression from multiple genes,and integration of metabolite feedback for expression control. Engineered trans-acting RNAs are flexible tools for rapid and specific control of gene expression. Naturally occurring bacterial small regulatory RNAs (sRNAs) can simultaneously target multiple mRNA transcripts via specific base-pairing, and can either stabilize or destabilize target transcripts. The importance and activity of these native small nucleic acids have been established, and it is time for their development for applications. The goal of the research is to create a ligand-responsive riboswitch-sRNA fusion molecule for improved fermentation cultures that produce biofuels and medicines. The Engineering of multi-target sRNA will enable coordinate and discoordinate mRNA stability/ translation control for metabolic engineering. Riboswitches integrated for control of RS-sRNAs will create a platform for user-specified targeting of mRNAs and integration with riboswitchbound metabolite feedback. Riboswitches integrated for stability control of sRNA will probe sRNA structural dynamics and activity, and will inform future design considerations for engineered sRNAs in their biological context.The proposed work will create fundamental tools for synthetic circuit development from the sRNAs, advance biochemical engineering by further bridging the disciplines of engineering and genetics, and enable us to train the next generation of biochemical engineers. The construction and analysis of modified sRNA genes can be parsed into smaller project subunits amenable to inclusion of undergraduates and summer students.

重新编程细菌以执行有用任务的能力建立在可以重新利用的现有遗传电路元件的基础上。合成生物电路设计面临的两个挑战包括多基因表达的协调，以及整合代谢物反馈以控制表达。工程反式作用rna是快速和特异性控制基因表达的灵活工具。天然存在的细菌小调控rna （sRNAs）可以通过特定的碱基配对同时靶向多个mRNA转录物，并且可以稳定或破坏目标转录物。这些天然小核酸的重要性和活性已经确立，现在是开发应用的时候了。这项研究的目标是创造一种配体反应性核糖体开关- srna融合分子，用于改善生产生物燃料和药物的发酵培养。多靶点sRNA的工程化将为代谢工程提供协调和不协调的mRNA稳定性/翻译控制。集成用于控制RS-sRNAs的核糖开关将为用户指定的mrna靶向和与核糖开关结合的代谢物反馈集成创建一个平台。集成用于sRNA稳定性控制的核糖开关将探测sRNA的结构动力学和活性，并将在其生物学背景下为工程sRNA的未来设计考虑提供信息。这项工作将为srna合成电路的开发创造基本工具，通过进一步连接工程和遗传学学科来推进生化工程，并使我们能够培养下一代生化工程师。修饰的sRNA基因的构建和分析可以被解析成更小的项目亚单位，适合本科生和暑期学生。