UNS: A Synthetic Bacterial Riboswitch Sensor for Microcystin Detection and Remediation

UNS：用于微囊藻毒素检测和修复的合成细菌核糖开关传感器

• 批准号: 1509022
• 负责人: David Wendell
• 金额: $ 29.99万
• 依托单位: University of Cincinnati Main Campus
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1509022&HistoricalAwards=false
• 关键词: UNS; Synthetic; Bacterial; Riboswitch; Sensor

1509022 Wendell, David Source water increasingly contains toxins produced by algal bloom. Easy and effective measurements of the toxins at concentrations that are toxic are not available. The researcher will develop a recombinant cell that will visually indicate if the toxins are present in source water. Successful completion of this project has the potential to reduce hazards associated with algal toxins. The toxins produced by cyanobacteria affecting human and animal health. The prevalence and toxicity of microcystin drives the need to create a sensor that is both affordable and highly sensitive, with direct applicability to environmental samples. Ribozymes, self-cleaving tertiary aggregates of RNA, have gained applications in gene expression. When RNA aptamers are incorporated into self-cleaving nucleic acid sequences, riboswitches responding solely to a desired ligand can be created. Using this strategy, riboswitch biosensors can be optimized to indicate the presence or absence of the toxin. The long term goal of this research is to produce a riboswitch-mediated biosensor orthogonal to the cyanobacterial toxin microcystin-LR. This bacterial sensor will optically report the presence of the toxin in the form of a simple color change and further assist with microcystin bioremediation by initializing a gene for microcystin-LR degradation (mlrA). The intellectual merit of this project is the design and construction of a riboswitch sensor that endows E. coli with a means of controlling gene expression in response to the presence of microcystin-LR. The central hypothesis is that a microcystin-LR specific riboswitch can predictably indicate the presence of microcystin-LR at very low, physiologically relevant concentrations. The basis for this hypothesis is preliminary results with a compendium of various ribosensor designs and a demonstrated response within the range of the World Health Organization required minimum.The broader impact of this project will be a cyanotoxin biosensor and bioremediation system that can be integrated into current potable water treatment systems and readily deployed in the field. In addition, this project will provide for the support and training of a graduate student, allow for the public presentation of the work at both academic conferences and a local high school, as well as recruitment of 3 underrepresented graduate and undergraduate students to environmental engineering. The 2 undergraduate recruits will be competing in the international synthetic biology competition iGEM, allowing them to build and share their genetically engineered projects with the global iGEM community.

1509022 温德尔，大卫源水越来越多地含有藻类水华产生的毒素。在有毒浓度下对毒素进行简单有效的测量是不可能的。研究人员将开发一种重组细胞，可以直观地指示水源水中是否存在毒素。 该项目的成功完成有可能减少与藻类毒素有关的危害。蓝细菌产生的毒素影响人类和动物的健康。微囊藻毒素的流行和毒性驱动了创建既负担得起又高度灵敏的传感器的需求，该传感器直接适用于环境样品。 核酶是一种自切割的RNA三级聚集体，在基因表达中有着广泛的应用。 当将RNA适体掺入自切割核酸序列中时，可以产生仅响应于所需配体的核糖开关。使用这种策略，可以优化核糖开关生物传感器，以指示毒素的存在或不存在。 本研究的长期目标是制作一种核糖开关介导的生物传感器正交的蓝藻毒素微囊藻毒素-LR。这种细菌传感器将以简单的颜色变化的形式光学报告毒素的存在，并通过初始化微囊藻毒素-LR降解基因（mlrA）进一步协助微囊藻毒素生物修复。该项目的智力价值是设计和构建核糖开关传感器，赋予E。大肠杆菌中的微囊藻毒素-LR的存在下，控制基因表达的手段。中心假设是微囊藻毒素-LR特异性核糖开关可以预测地指示微囊藻毒素-LR在非常低的生理相关浓度下的存在。这一假设的基础是初步的结果与纲要的各种ribosensor的设计和世界卫生组织所需的范围内的响应minimal.The更广泛的影响，这个项目将是一个氰毒素生物传感器和生物修复系统，可以集成到目前的饮用水处理系统，并随时部署在外地。 此外，该项目还将为一名研究生提供支助和培训，以便在学术会议和当地一所高中公开介绍其工作，并招聘3名代表性不足的研究生和本科生从事环境工程工作。 这两名本科生将参加国际合成生物学竞赛iGEM，让他们能够与全球iGEM社区建立和分享他们的基因工程项目。