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A cis-acting ligase ribozyme generates circular RNA in vitro for ectopic protein functioning

Author

Listed:
  • Chan-I Su

    (National Health Research Institutes)

  • Zih-Shiuan Chuang

    (National Health Research Institutes
    National Health Research Institutes)

  • Chi-Ting Shie

    (National Health Research Institutes)

  • Hsin-I Wang

    (National Health Research Institutes)

  • Yu-Ting Kao

    (National Health Research Institutes)

  • Chia-Yi Yu

    (National Health Research Institutes
    National Health Research Institutes
    National Cheng Kung University)

Abstract

Delivering synthetic protein-coding RNA bypassing the DNA stage for ectopic protein functioning is a novel therapeutic strategy. Joining the linear RNA head-to-tail covalently could be a state-of-the-art strategy for functioning longer. Here we enroll a cis-acting ligase ribozyme (RzL) to generate circular RNA (circRNA) in vitro for ectopic protein expression. The RNA circularization is confirmed by masking the 5’ phosphate group, resisting exonuclease RNase R digestion, failing for further tailing, and sequencing the RT-PCR products of the joined region. Interestingly, one internal ribosome entry site (IRES) renders circRNA translation competent, but two IRES in cis, not trans, hamper the translation. The circRNA with highly potent in translation is conferred for antiviral functioning. Accompanying specific guided RNA, a circRNA expressing ribonuclease Cas13 shows excellent potential against the corresponding RNA virus, further extending circRNA functioning in its growing list of applications.

Suggested Citation

  • Chan-I Su & Zih-Shiuan Chuang & Chi-Ting Shie & Hsin-I Wang & Yu-Ting Kao & Chia-Yi Yu, 2024. "A cis-acting ligase ribozyme generates circular RNA in vitro for ectopic protein functioning," Nature Communications, Nature, vol. 15(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-51044-y
    DOI: 10.1038/s41467-024-51044-y
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    References listed on IDEAS

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    1. R. Alexander Wesselhoeft & Piotr S. Kowalski & Daniel G. Anderson, 2018. "Engineering circular RNA for potent and stable translation in eukaryotic cells," Nature Communications, Nature, vol. 9(1), pages 1-10, December.
    2. Florian Krammer, 2020. "SARS-CoV-2 vaccines in development," Nature, Nature, vol. 586(7830), pages 516-527, October.
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