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

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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.