PT  - JOURNAL ARTICLE
AU  - Alam, Khalid K.
AU  - Jung, Jaeyoung K.
AU  - Verosloff, Matthew S.
AU  - Clauer, Phillip R.
AU  - Lee, Jeong Wook
AU  - Capdevila, Daiana A.
AU  - Pastén, Pablo A.
AU  - Giedroc, David P.
AU  - Collins, James J.
AU  - Lucks, Julius B.
TI  - Rapid, Low-Cost Detection of Water Contaminants Using Regulated <em>In Vitro</em> Transcription
AID  - 10.1101/619296
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 619296
4099  - http://biorxiv.org/content/early/2019/05/02/619296.short
4100  - http://biorxiv.org/content/early/2019/05/02/619296.full
AB  - Synthetic biology has enabled the development of powerful nucleic acid diagnostic technologies for detecting pathogens and human health biomarkers. Here we expand the reach of synthetic biology-enabled diagnostics by developing a cell-free biosensing platform that uses RNA output sensors activated by ligand induction (ROSALIND) to detect harmful contaminants in aqueous samples. ROSALIND consists of three programmable components: highly-processive RNA polymerases, allosteric transcription factors, and synthetic DNA transcription templates. Together, these components allosterically regulate the in vitro transcription of a fluorescence-activating RNA aptamer: in the absence of a target compound, transcription is blocked, while in its presence a fluorescent signal is produced. We demonstrate that ROSALIND can be configured to detect a range of water contaminants, including antibiotics, toxic small molecules, and metals. Our cell-free biosensing platform, which can be freeze-dried for field deployment, creates a new capability for point-of-use monitoring of molecular species to address growing global crises in water quality and human health.