Engineering circular RNA for potent and stable translation in eukaryotic cells

By A. Wesselhoeft, Piotr S. Kowalski, Daniel G. Anderson. Nature Communications, 2018

706 citations recorded by the source corpus.

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. Circular RNAs have recently been shown to have protein-coding potential. Here the authors design a self-splicing RNA that, when circularized, provides for stable high-yield protein production.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/1a4ee7ac0868ac371710e8c1019ff497f6d8b677

Citations in the corpus

• Circular RNA-protein interactions: functions, mechanisms, and identification (2020, background): “Although the physiological functions of most of these proteins have not yet been identified, it is likely that they share some of the functional abilities of their full-length protein counterparts encoded by the linear forms of the transcripts [108].”
• Past, present, and future of circRNAs (2019, critical): “However, the magnitude of RIG-1 activation due to exogenous circRNA was challenged by later reports (Wesselhoeft et al , 2018, 2019).”