Engineering circular RNA for enhanced protein production

Publication Date: 17 August 2022

Chen, R. et al. (2022). Engineering circular RNA for enhanced protein production. Nat. Biotechnol. DOI: 10.1038/s41587-022-01393-0

The success of mRNA vaccines has prompted interest in extending the duration of protein expression for gene therapy applications. Circular RNAs (circRNAs), in which coding RNA molecules are joined head to tail, show significant promise in this regard.

In this study, the researchers report the development of a modular high-throughput platform to make and test synthetic circRNAs. To maximize circRNA translation, they optimized five elements: vector topology, 5′ and 3′ untranslated regions, internal ribosome entry sites and synthetic aptamers recruiting translation initiation machinery. To optimize translation activity, they prepared NanoLuc® luciferase reporter constructs and transfected HeLa cells. They also examined circRNA expression levels in vivo by luminescence imaging in mice. The authors conclude that their optimized circRNA synthesis platform increased circRNA protein yields by several hundred-fold and enabled potent and durable protein production in vivo.

Keywords: gene delivery, gene therapy, nucleic acid therapeutics, RNA splicing