Engineering circular RNA for potent and stable translation in eukaryotic cells

R. Alexander Wesselhoeft, Piotr S. Kowalski and Daniel G. Anderson ()
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R. Alexander Wesselhoeft: Massachusetts Institute of Technology
Piotr S. Kowalski: Massachusetts Institute of Technology
Daniel G. Anderson: Massachusetts Institute of Technology

Nature Communications, 2018, vol. 9, issue 1, 1-10

Abstract: Abstract Messenger RNA (mRNA) has broad potential for application in biological systems. However, one fundamental limitation to its use is its relatively short half-life in biological systems. Here we develop exogenous circular RNA (circRNA) to extend the duration of protein expression from full-length RNA messages. First, we engineer a self-splicing intron to efficiently circularize a wide range of RNAs up to 5 kb in length in vitro by rationally designing ubiquitous accessory sequences that aid in splicing. We maximize translation of functional protein from these circRNAs in eukaryotic cells, and we find that engineered circRNA purified by high performance liquid chromatography displays exceptional protein production qualities in terms of both quantity of protein produced and stability of production. This study pioneers the use of exogenous circRNA for robust and stable protein expression in eukaryotic cells and demonstrates that circRNA is a promising alternative to linear mRNA.

Date: 2018
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DOI: 10.1038/s41467-018-05096-6

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