TY - JOUR
T1 - Engineering circular RNA for enhanced protein production
AU - Chen, Robert
AU - Wang, Sean K.
AU - Belk, Julia A.
AU - Amaya, Laura
AU - Li, Zhijian
AU - Cardenas, Angel
AU - Abe, Brian T.
AU - Chen, Chun Kan
AU - Wender, Paul A.
AU - Chang, Howard Y.
N1 - Publisher Copyright:
© 2022, The Author(s).
PY - 2023/2
Y1 - 2023/2
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85134521147&partnerID=8YFLogxK
U2 - 10.1038/s41587-022-01393-0
DO - 10.1038/s41587-022-01393-0
M3 - Article
C2 - 35851375
AN - SCOPUS:85134521147
SN - 1087-0156
VL - 41
SP - 262
EP - 272
JO - Nature Biotechnology
JF - Nature Biotechnology
IS - 2
ER -