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  3. Engineering circular RNA for potent and stable translation in eukaryotic cells

Engineering circular RNA for potent and stable translation in eukaryotic cells

  • Nat Commun. 2018 Jul 6;9(1):2629. doi: 10.1038/s41467-018-05096-6.
R Alexander Wesselhoeft 1 2,  Piotr S Kowalski 3,  Daniel G Anderson 4 5 6 7
Affiliations

Affiliations

  • 1 David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
  • 2 Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
  • 3 Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
  • 4 David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. dgander@mit.edu.
  • 5 Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. dgander@mit.edu.
  • 6 Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. dgander@mit.edu.
  • 7 Harvard and MIT Division of Health Science and Technology, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. dgander@mit.edu.
PMID: 29980667 DOI: 10.1038/s41467-018-05096-6
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.

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