Engineering circular RNA for enhanced protein production

Circular RNAs (circRNAs) are stable and prevalent RNAs in eukaryotic cells that arise from back-splicing. Synthetic circRNAs and some endogenous circRNAs can encode proteins, raising the promise of circRNA as a platform for gene expression. In this study, we developed a systematic approach for rapid assembly and testing of features that affect protein production from synthetic circRNAs. To maximize circRNA translation, we optimized five elements: vector topology, 5' and 3' untranslated regions, internal ribosome entry sites and synthetic aptamers recruiting translation initiation machinery. Together, these design principles improve circRNA protein yields by several hundred-fold, provide increased translation…

• NS
  National Science Foundation

  Awards: DGE-1656518, -1656518, 1656518, 1856414
• HH
  Howard Hughes Medical Institute
• NI
  National Institutes of Health

  Awards: R35-CA209919, CA031845, NIH R01, 5T32GM008412
• DO
  Division of Chemistry

  Award: CHE-1856414