Engineering circular RNA for enhanced protein production.

Chen, Robert; Wang, Sean K; Belk, Julia A; Amaya, Laura; Li, Zhijian; Cardenas, Angel; Abe, Brian T; Chen, Chun-Kan et al. · Nat Biotechnol · 2023

basic_science · Level V

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Abstract

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 over messenger RNA in vitro, provide more durable translation in vivo and are generalizable across multiple transgenes.

Medical subject headings