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

1509022大卫温德尔水源水越来越多地含有由藻类水华产生的毒素。在有毒的浓度下，没有简单有效的毒素测量方法。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险。蓝藻产生的影响人类和动物健康的毒素。微囊藻毒素的流行和毒性促使人们需要创造一种既负担得起又高度灵敏的传感器，直接适用于环境样品。核酶是一种自切割的RNA三聚体，已在基因表达中得到应用。当RNA适配子被加入到自切割的核酸序列中时，就可以产生只对所需配体反应的核糖开关。使用这一策略，核糖开关生物传感器可以被优化以指示毒素的存在或不存在。本研究的长期目标是制造一种与蓝藻毒素微囊藻毒素-LR垂直的核糖开关介导的生物传感器。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)进一步辅助微囊藻毒素的生物修复。这个项目的智力价值在于设计和建造了一种核糖开关传感器，使大肠杆菌能够控制基因表达，以应对微囊藻毒素-LR的存在。中心假设是，微囊藻毒素-LR特异性核糖开关可以预测在非常低的生理相关浓度下微囊藻毒素-LR的存在。这一假设的基础是各种核糖传感器设计概要的初步结果，以及在世界卫生组织要求的最低限度范围内证明的响应。该项目的更广泛影响将是一种氰毒素生物传感器和生物修复系统，可以集成到当前的饮用水处理系统中，并易于在现场部署。此外，该项目将为一名研究生提供支助和培训，允许在学术会议和当地一所高中公开介绍这项工作，并为环境工程学招收3名代表性不足的研究生和本科生。这两名本科生将参加国际合成生物学大赛iGEM，使他们能够构建并与全球iGEM社区分享他们的基因工程项目。

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.