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<p>Ribozyme-based approaches have been extensively employed for in vitro circular RNA (circRNA) synthesis. However, several critical challenges are impeding the progress in this field, including the relatively low ribozyme utilization efficiency, limited synthetic flexibility, and complicated downstream purification procedures. Here, we introduce a split intron-exon (SIE) system for circRNA synthesis by physically separating the ribozyme from its substrates. This design ensures efficient circRNA synthesis by overcoming the inherent limitations of current ribozyme-based techniques, in which each ribozyme can only catalyse the synthesis of at most one circRNA molecule. Moreover, the SIE system enables the efficient synthesis of both unmodified and chemically modified circRNAs. Furthermore, the recyclability and immobilization of the ribozyme within the SIE system reduce reaction impurities and improves circRNA enrichment and yield. Overall, the SIE system holds great potential to advance the diversification and large-scale development of circRNA therapeutics.</p>

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Split intron-exon system for circular RNA synthesis

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年7月28日

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  • Lei Wang,
  • Qiaoli Zhai,
  • Chunbo Dong,
  • Weibing Zhang,
  • Xu Ma,
  • Kai Yang,
  • Shaolong Qi,
  • Yarong Wu,
  • Guocan Yu,
  • Lian-Qing Wang,
  • George Fu Gao,
  • Zhida Liu

摘要

Ribozyme-based approaches have been extensively employed for in vitro circular RNA (circRNA) synthesis. However, several critical challenges are impeding the progress in this field, including the relatively low ribozyme utilization efficiency, limited synthetic flexibility, and complicated downstream purification procedures. Here, we introduce a split intron-exon (SIE) system for circRNA synthesis by physically separating the ribozyme from its substrates. This design ensures efficient circRNA synthesis by overcoming the inherent limitations of current ribozyme-based techniques, in which each ribozyme can only catalyse the synthesis of at most one circRNA molecule. Moreover, the SIE system enables the efficient synthesis of both unmodified and chemically modified circRNAs. Furthermore, the recyclability and immobilization of the ribozyme within the SIE system reduce reaction impurities and improves circRNA enrichment and yield. Overall, the SIE system holds great potential to advance the diversification and large-scale development of circRNA therapeutics.