Abstract
ROSALIND (RNA Output Sensors Activated by Ligand Induction) is an in vitro biosensing system that detects small molecules using regulated transcription reactions. It consists of three key components: (1) RNA polymerases, (2) allosteric protein transcription factors, and (3) synthetic DNA transcription templates that together regulate the synthesis of a fluorescence-activating RNA aptamer. The system can detect a wide range of chemicals including antibiotics, small molecules, and metal ions. We have demonstrated that ROSALIND can be lyophilized and transported at ambient conditions for water testing on-site. Here, we describe how to set up a ROSALIND reaction for detecting various chemical contaminants in water using a model transcription factor as well as how to build a new ROSALIND sensor.
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Acknowledgments
This work was supported by NSF MCB RAPID (grant no. 1929912 to J.B.L.), the Crown Family Center for Jewish and Israel Studies at Northwestern University (to J.B.L.), Searle Funds at the Chicago Community Trust (to J.B.L.) and a Ryan Fellowship at Northwestern University (to J.K.J.).
Competing Interests
J.K.J., K.K.A. and J.B.L. have submitted a US provisional patent application (no. 62/758,242) relating to regulated IVT reactions and a US provisional patent application (no. 62/838,852) relating to the preservation and stabilization of IVT reactions. K.K.A. and J.B.L. are founders and have a financial interest in Stemloop, Inc. The latter interests are reviewed and managed by Northwestern University in accordance with their conflict-of-interest policies.
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Jung, J.K., Alam, K.K., Lucks, J.B. (2022). ROSALIND: Rapid Detection of Chemical Contaminants with In Vitro Transcription Factor-Based Biosensors. In: Karim, A.S., Jewett, M.C. (eds) Cell-Free Gene Expression. Methods in Molecular Biology, vol 2433. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1998-8_20
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DOI: https://doi.org/10.1007/978-1-0716-1998-8_20
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