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...

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Bibliographic Details
Published inNature biotechnology Vol. 41; no. 2; pp. 262 - 272
Main Authors Chen, Robert, Wang, Sean K., Belk, Julia A., Amaya, Laura, Li, Zhijian, Cardenas, Angel, Abe, Brian T., Chen, Chun-Kan, Wender, Paul A., Chang, Howard Y.
Format Journal Article
LanguageEnglish
Published New York Nature Publishing Group US 01.02.2023
Nature Publishing Group
Subjects
3' Untranslated regions
631/1647/2300
631/208/1792
631/61/201
631/61/51/391
Agriculture
Aptamers
Bioinformatics
Biomedical and Life Sciences
Biomedical Engineering/Biotechnology
Biomedicine
Biotechnology
Circular RNA
Cloning
Design optimization
Gene expression
In vivo methods and tests
Life Sciences
mRNA
Protein folding
Proteins
Ribonucleic acid
RNA
RNA - genetics
RNA - metabolism
RNA Splicing
RNA, Circular - genetics
RNA, Messenger - genetics
RNA, Messenger - metabolism
Severe acute respiratory syndrome coronavirus 2
Splicing
Topology
Transgenes
Translation
Translation initiation
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Summary: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. Protein expression from circular RNAs is enhanced several hundred-fold by optimizing vector design.
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ISSN:1087-0156
1546-1696
1546-1696
DOI:10.1038/s41587-022-01393-0