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Production and Purification of Artificial Circular RNA Sponges for Application in Molecular Biology and Medicine

Institute of Biochemistry, Faculty of Biology and Chemistry, University of Giessen, Heinrich-Buff-Ring 17, D-35392 Giessen, Germany
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Academic Editor: Emanuela Jacchetti
Methods Protoc. 2020, 3(2), 42; https://doi.org/10.3390/mps3020042
Received: 27 April 2020 / Revised: 21 May 2020 / Accepted: 22 May 2020 / Published: 26 May 2020
Characterized by their covalently closed structure and thus an elevated stability compared to linear RNA molecules, circular RNAs (circRNAs) form a novel class of mainly non-coding RNAs. Although the biological functions of naturally occurring circRNAs are largely unknown, they were reported to act as molecular sponges, sequestering microRNAs (miRNAs), resulting in a de-repression of target mRNAs. Taking these characteristics of naturally occurring circRNAs into account, artificial circRNAs could be a potential tool in molecular biology and medicine. Using the Hepatitis C virus (HCV) as a model system, this application of artificial circular RNAs was demonstrated. The virus requires cellular miRNA miR-122 for its life cycle, and circRNAs specifically engineered to efficiently sequester this miRNA impacted viral propagation. Since in this context the production of engineered circRNA remains the limiting factor, we present a method to produce and efficiently purify artificial circRNA sponges (ciRS) in vitro. In this protocol we provide insights into a small-scale and large-scale production technique of artificial circular RNA sponges relying on in vitro transcription and RNA ligation. View Full-Text
Keywords: circular RNA; circular RNA sponges; microRNA; RNA therapy; molecular sponge; circRNA production; artificial circular RNA sponge; artificial circRNA; ciRS circular RNA; circular RNA sponges; microRNA; RNA therapy; molecular sponge; circRNA production; artificial circular RNA sponge; artificial circRNA; ciRS
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MDPI and ACS Style

Breuer, J.; Rossbach, O. Production and Purification of Artificial Circular RNA Sponges for Application in Molecular Biology and Medicine. Methods Protoc. 2020, 3, 42. https://doi.org/10.3390/mps3020042

AMA Style

Breuer J, Rossbach O. Production and Purification of Artificial Circular RNA Sponges for Application in Molecular Biology and Medicine. Methods and Protocols. 2020; 3(2):42. https://doi.org/10.3390/mps3020042

Chicago/Turabian Style

Breuer, Janina, and Oliver Rossbach. 2020. "Production and Purification of Artificial Circular RNA Sponges for Application in Molecular Biology and Medicine" Methods and Protocols 3, no. 2: 42. https://doi.org/10.3390/mps3020042

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