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

水源水中越来越多地含有由藻华产生的毒素。目前还没有对有毒浓度的毒素进行简单而有效的测量。研究人员将开发一种重组细胞，可以直观地显示水源中是否存在毒素。这个项目的成功完成有可能减少与藻类毒素有关的危害。蓝藻产生的毒素影响人类和动物的健康。微囊藻毒素的普遍性和毒性促使人们需要创造一种既负担得起又高度敏感的传感器，并直接适用于环境样品。核糖酶是RNA的自切割三级聚集体，在基因表达中得到了应用。当RNA适体被整合到自切割核酸序列中时，可以创建仅响应所需配体的核开关。使用这种策略，核糖开关生物传感器可以优化，以指示毒素的存在或不存在。本研究的长期目标是生产一种与蓝藻毒素微囊藻毒素- lr正交的核糖体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在光学上报告毒素的存在，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。本项目的智力优势在于设计和构建了一个核糖开关传感器，该传感器赋予大肠杆菌一种控制基因表达的手段，以应对微囊藻毒素- lr的存在。核心假设是，微囊藻毒素- lr特异性核糖体开关可以预测微囊藻毒素- lr在非常低的生理相关浓度下的存在。这一假设的基础是各种核传感器设计的概要的初步结果，以及在世界卫生组织要求的最低限度范围内证明的反应。该项目的广泛影响将是一个蓝藻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，并随时部署在现场。此外，该项目将为一名研究生提供支持和培训，允许在学术会议和一所当地高中公开介绍工作，并招募3名代表性不足的研究生和本科生学习环境工程。这两名本科生新兵将参加国际合成生物学竞赛iGEM，使他们能够建立并与全球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.