Journal
NATURE BIOTECHNOLOGY
Volume 38, Issue 12, Pages -Publisher
NATURE PORTFOLIO
DOI: 10.1038/s41587-020-0571-7
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Funding
- Northwestern University's Graduate School Cluster in Biotechnology, System, and Synthetic Biology
- National Science Foundation (NSF) Synthetic Biology REU [DBI-1757973]
- Pew Charitable Trusts
- National Institutes of Health [R35 GM118157]
- CONICYT/FONDECYT [1161337]
- ANID/FONDAP [15110020]
- NSF [1452441, 1929912]
- Crown Family Center for Jewish and Israel Studies at Northwestern University
- Searle Funds at the Chicago Community Trust
- Direct For Biological Sciences
- Div Of Molecular and Cellular Bioscience [1929912] Funding Source: National Science Foundation
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Lack of access to safe drinking water is a global problem, and methods to reliably and easily detect contaminants could be transformative. We report the development of a cell-free in vitro transcription system that uses RNA Output Sensors Activated by Ligand Induction (ROSALIND) to detect contaminants in water. A combination of highly processive RNA polymerases, allosteric protein transcription factors and synthetic DNA transcription templates regulates the synthesis of a fluorescence-activating RNA aptamer. The presence of a target contaminant induces the transcription of the aptamer, and a fluorescent signal is produced. We apply ROSALIND to detect a range of water contaminants, including antibiotics, small molecules and metals. We also show that adding RNA circuitry can invert responses, reduce crosstalk and improve sensitivity without protein engineering. The ROSALIND system can be freeze-dried for easy storage and distribution, and we apply it in the field to test municipal water supplies, demonstrating its potential use for monitoring water quality.
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