CAREER: Waterborne Cyanotoxin Detection and Removal Using DNA-Based Affinity Reagents

1552961 HeemstraThe总体目标这个NSF职业生涯的建议是开发新的方法检测和去除水传播的蓝藻毒素，并显着增加学生的研究经验的访问。蓝藻水华发生在淡水湖泊和水库作为环境条件的结果，导致小分子蓝藻毒素释放到水中。这些毒素对人类健康构成严重威胁，接触这些毒素会导致急性和慢性疾病。蓝藻污染尤其是磷和氮燃料蓝藻生长促进的水源加速富营养化的问题，正如在2015年伊利湖事件中观察到的那样。拟议的研究和推广活动建立在首席研究员（PI）建立的坚实而成功的基础上。PI开发了用于可卡因的DNA分裂aptmer技术，并证明了她的团队使用该技术以三明治形式构建侧向流传感器的能力。PI将应用这些知识创建DNA裂解适体，用于检测两种蓝藻毒素-微囊藻毒素和石房蛤毒素。发达国家积极监测饮用水中的藻类毒素水平。然而，全球大部分饮用水供应缺乏这种监测。毒素暴露经常是由于娱乐用水造成的，而广泛的监测是不切实际的。其中最普遍和有害的这些水载小分子毒素是微囊藻毒素-LR和石房蛤毒素。拟议的研究活动探索了两个新的、潜在的变革性概念：（1）使用DNA适体功能化膜选择性地从饮用水中去除毒素，以及（2）应用DNA分裂适体连接来构建廉价、用户友好的侧向流传感器。除了产生新技术外，这项研究还将深入了解DNA组装和DNA-小分子相互作用，并将建立一种可靠有效的方法来产生DNA分裂适体。这将为旨在开发检测和去除小分子水污染物的新方法的长期努力奠定基础。此外，拟议活动的未来扩展可侧重于开发一个用户界面，使公众能够报告水传播毒素的阳性检测结果。这将利用众包的力量在公众中分享有关污染事件的信息，并为科学家和工程师提供数据，帮助他们更好地了解导致有毒蓝藻水华的环境条件。

1552961HeemstraThe overarching goals of this NSF CAREER proposal are to develop new methods for the detection and removal of waterborne cyanotoxins and to dramatically increase the access of students to research experiences. Cyanobacterial blooms occur in freshwater lakes and reservoirs as a result of environmental conditions, resulting in the release of small-molecule cyanotoxins into the water. These toxins pose a serious threat to human health, with exposure resulting in both acute and chronic disease. Cyanotoxin contamination is especially problematic with the accelerated eutrophication of water sources promoted by phosphorus- and nitrogen-fueled cyanobacterial growth, as was observed in the Lake Erie incidence in 2015.The proposed research and outreach activities build from a firm and successful foundation established by the principal investigator (PI). The PI developed DNA split aptmer technology for cocaine and has demonstrated her group's ability to construct lateral flow sensors using this technology in a sandwich format. The PI will apply this knowledge to create DNA split aptmers for the detection of two cyanotoxins - microcystin and saxitoxin. Algal toxin levels in drinking water are actively monitored in developed nations. However, such monitoring is lacking for much of the global drinking water supply. Toxin exposure frequently results from recreational water use where widespread monitoring is impractical. Among the most prevalent and harmful of these waterborne small-molecule toxins are microcystin-LR and saxitoxin. The proposed research activities explore two novel, and potentially transformative concepts: (1) The use of DNA aptamer-functionalized membranes to selectively remove toxins from drinking water, and (2) The application of DNA split aptamer ligation to construct inexpensive, user-friendly lateral flow sensors. In addition to generating new technologies, this research will provide insight into DNA assembly and DNA-small molecule interactions, and will establish a reliable and efficient method for generating DNA split aptamers. This will lay a foundation for a long-term effort aimed at developing new methods for the detection and removal of small-molecule water contaminants. Additionally, future expansion of the proposed activities could focus on developing a user interface that would enable the general public to report positive test results for waterborne toxins. This would harness the power of crowdsourcing to share information among the public regarding contamination events, and would provide scientists and engineers with data to help them better understand the environmental conditions that give rise to toxic cyanobacterial blooms.