Engineering Circular RNA for Enhanced Protein Production

Overview

Journal Nat Biotechnol

Specialty Biotechnology

Date 2022 Jul 19

PMID 35851375

Authors

Robert Chen

Sean K Wang

Julia A Belk

Laura Amaya

Zhijian Li

Angel Cardenas

Brian T Abe

Chun-Kan Chen

Paul A Wender

Howard Y Chang

Affiliations

Soon will be listed here.

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.

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