Engineering circular RNA for potent and stable translation in eukaryotic cells
Name
s41467-018-05096-6.pdf
Size
1.8 MB
Format
Adobe PDF
Checksum (MD5)
bd918d9a68fc9b0246cb9cb1e7fada08
Author(s)
Wesselhoeft IV, Robert Alexander
Kowalski, Piotr S
Anderson, Daniel Griffith
Date Issued
July 2018
Journal
Nature Communications
Publisher
Nature Publishing Group
Citation
Wesselhoeft, R. Alexander, Piotr S. Kowalski, and Daniel G. Anderson. “Engineering Circular RNA for Potent and Stable Translation in Eukaryotic Cells.” Nature Communications 9, no. 1 (July 6, 2018).
Version
Final published version
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.
MIT Department
Harvard University--MIT Division of Health Sciences and Technology
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Chemical Engineering
Koch Institute for Integrative Cancer Research at MIT
Terms of Use
Creative Commons Attribution 4.0 International License
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1038/s41467-018-05096-6
