CircRNA是一种参与多种生物学功能和疾病的重要RNA分子，通常在肿瘤组织中表现出表达降低，发挥肿瘤抑制因子的作用，并提示其具有治疗癌症的潜力。然而，目前促进circRNA产生的方法是有限的。这项研究介绍了一种增强 circRNA 生物合成的新方法，称为 circRNA 促进 RNA (cpRNA)。 CpRNA 旨在补充前体 mRNA 内反向互补匹配 (RCM) 的侧翼序列，从而通过改善外显子环化促进 circRNA 形成。使用 split-GFP 报告系统，我们证明 cpRNA 显着增强 circGFP 的产生。优化确定了 cpRNA 促进 circRNA 生物发生的最佳条件，然后使用这些 cpRNA 来增加内源 circRNA 的产生。这些结果表明，cpRNA可以特异性增加癌细胞中RCM的内源性circRNA的产生，例如circZKSCAN1和circSMARCA5，从而通过调节circRNA相关途径抑制细胞增殖和迁移，展示了cpRNA的治疗潜力。机制研究还表明，cpRNA 部分通过拮抗 DHX9 的解旋功能来促进 circRNA 生物发生。总体而言，这些发现表明 cpRNA 代表了 circRNA 过度表达的一种有前景的策略，为以低 circRNA 水平为标志的疾病提供了潜在的治疗方法。© 作者 2024。由牛津大学出版社代表 Nucleic Acids Research 出版。

CircRNA, an essential RNA molecule involved in various biological functions and diseases, often exhibits decreased expression in tumor tissues, playing a role as a tumor suppressor, and suggesting therapeutic potential for cancer. However, current methods for promoting circRNA production are limited. This study introduces a novel approach for enhancing circRNA biogenesis, termed circRNA promoting RNA (cpRNA). CpRNA is designed to complement the flanking sequences of reverse complementary matches (RCMs) within pre-mRNA, thereby facilitating circRNA formation through improved exon circularization. Using a split-GFP reporter system, we demonstrated that cpRNA significantly enhance circGFP production. Optimization identified the best conditions for cpRNA to promote circRNA biogenesis, and these cpRNAs were then used to augment the production of endogenous circRNAs. These results indicate that cpRNAs can specifically increase the production of endogenous circRNAs with RCMs, such as circZKSCAN1 and circSMARCA5 in cancer cells, thereby inhibiting cell proliferation and migration by modulating circRNA-related pathways, showcasing the therapeutic potential of cpRNAs. Mechanistic studies have also shown that cpRNA promotes circRNA biogenesis, in part, by antagonizing the unwinding function of DHX9. Overall, these findings suggest that cpRNA represents a promising strategy for circRNA overexpression, offering a potential treatment for diseases marked by low circRNA levels.© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.