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

1552961Heemstra 这项 NSF 职业提案的总体目标是开发检测和去除水性蓝藻毒素的新方法，并大幅增加学生获得研究经验的机会。由于环境条件，淡水湖泊和水库中会发生蓝藻水华，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。正如 2015 年伊利湖事件中所观察到的那样，以磷和氮为燃料的蓝藻生长加速了水源的富营养化，蓝藻毒素污染问题尤其严重。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之上。该 PI 开发了用于可卡因的 DNA 分裂适体技术，并展示了她的团队使用该技术以三明治形式构建侧向流动传感器的能力。 PI 将应用这些知识来创建 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都缺乏这种监测。毒素暴露常常是由于娱乐用水造成的，而广泛的监测是不切实际的。这些水性小分子毒素中最普遍和最有害的是微囊藻毒素-LR 和石房蛤毒素。拟议的研究活动探索了两个新颖且具有潜在变革性的概念：（1）使用DNA适体功能化膜选择性地去除饮用水中的毒素，以及（2）应用DNA分裂适体连接来构建廉价、用户友好的侧流传感器。除了产生新技术之外，这项研究还将深入了解 DNA 组装和 DNA-小分子相互作用，并将建立一种可靠且有效的方法来生成 DNA 裂解适体。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能会集中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之间共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大量繁殖的环境条件。

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.