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Molecules 2016, 21(10), 1310;

Real-Time Detection of a Self-Replicating RNA Enzyme

Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037, USA
Author to whom correspondence should be addressed.
Academic Editor: Sabine Müller
Received: 6 September 2016 / Revised: 25 September 2016 / Accepted: 27 September 2016 / Published: 30 September 2016
(This article belongs to the Special Issue Ribozymes and RNA Catalysis)
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A system was developed to detect the self-replication of an RNA enzyme in real time. The enzyme is an RNA ligase that undergoes exponential amplification at a constant temperature and can be made to operate in a ligand-dependent manner. The real-time system is based on a fluorimetric readout that directly couples the ligation event to an increase in florescence signal that can be monitored using standard instrumentation. The real-time system can also operate entirely with l-RNA, which is not susceptible to degradation by ribonucleases that are present in biological samples. The system is analogous to real-time PCR, but with the potential to detect small molecules, proteins, and other targets that can be recognized by a suitable aptamer. The ligand-dependent self-replication of RNA has potential applications in molecular diagnostics and biosensing that benefit from the rapid, precise, and real-time detection of various target molecules. View Full-Text
Keywords: aptazyme; exponential amplification; fluorescence detection; in vitro selection; RNA enzyme; RNA ligation; self-replication aptazyme; exponential amplification; fluorescence detection; in vitro selection; RNA enzyme; RNA ligation; self-replication

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Olea, C.; Joyce, G.F. Real-Time Detection of a Self-Replicating RNA Enzyme. Molecules 2016, 21, 1310.

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