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

1552961 heemstra1这个国家科学基金会职业建议的总体目标是开发检测和去除水生蓝藻毒素的新方法，并大大增加学生获得研究经验的机会。由于环境条件的影响，淡水湖和水库中会出现蓝藻繁殖，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，接触这些毒素可导致急性和慢性疾病。正如2015年伊利湖事件所观察到的那样，磷和氮驱动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）建立的坚实和成功的基础之上。PI开发了可卡因的DNA分裂适体技术，并展示了她的团队利用该技术在三明治格式中构建横向流量传感器的能力。PI将运用这些知识来创建DNA分裂核酸适体，用于检测两种蓝藻毒素——微囊藻毒素和蛤蚌毒素。发达国家积极监测饮用水中的藻类毒素水平。然而，对全球大部分饮用水供应缺乏这样的监测。毒素暴露往往是由于娱乐用水造成的，而广泛监测是不切实际的。在这些水传播的小分子毒素中，最普遍和最有害的是微囊藻毒素和蛤蚌毒素。我们提出的研究活动探索了两个新颖且具有潜在变革意义的概念：(1)使用DNA适配体功能化膜选择性地去除饮用水中的毒素；(2)应用DNA分裂适配体连接构建廉价、用户友好的横向流量传感器。除了产生新技术外，这项研究还将为DNA组装和DNA-小分子相互作用提供见解，并将建立一种可靠而有效的方法来产生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，今后扩大拟议的活动可侧重于开发一种用户界面，使一般公众能够报告水媒毒素的阳性检测结果。这将利用众包的力量，在公众之间分享有关污染事件的信息，并将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.