Militello Giuseppe, Greig Alyssa, Bi Chongfeng, Vasileva Ana, Zavodszky Maria I, Lo Shih-Ching, Guilmette Edward, Clarner Pete, Liu Bin, Bhat Guruharsha, Suh Junghae, Dow Lukas, Zuber Johannes, Fellmann Christof, Premsrirut Prem K
Mirimus Inc., 760 Parkside Avenue, Suite 206, Brooklyn, NY, 11226, USA.
SUNY Downstate Medical Center, 450 Clarkson Ave, Brooklyn, NY, 11203, USA.
Sci Rep. 2025 Jul 1;15(1):21419. doi: 10.1038/s41598-025-07061-y.
RNA interference (RNAi) is emerging as a powerful strategy for therapeutic targeting of "undruggable" targets. However, efficacy of currently used siRNA-based therapies is often hindered by transient effects and limited modeling possibilities. Artificial microRNAs (amiRNAs or miRNA scaffolds) present a durable and precise approach to gene silencing, opening new avenues for developing long lasting targeted therapies. In this study, we engineered highly expressed primary miRNAs (pri-miRNAs) with sequence determinants known to enhance processing efficacy and precision. The resulting amiRNAs were extensively tested both in vitro and in vivo and proved to efficiently silence a target gene when virally delivered via adeno-associated virus (AAV) into mice brains. This study provides a set of novel amiRNAs with potential therapeutic application as well as a pipeline to generate and validate novel amiRNAs from endogenous pri-miRNAs.
RNA干扰（RNAi）正成为一种针对“不可成药”靶点进行治疗性靶向的强大策略。然而，目前基于小干扰RNA（siRNA）的疗法的疗效常常受到短暂效应和有限建模可能性的阻碍。人工微小RNA（amiRNA或miRNA支架）为基因沉默提供了一种持久且精确的方法，为开发长效靶向疗法开辟了新途径。在本研究中，我们设计了具有已知可提高加工效率和精度的序列决定因素的高表达初级微小RNA（pri-miRNA）。所产生的amiRNA在体外和体内都进行了广泛测试，并且当通过腺相关病毒（AAV）病毒递送至小鼠大脑时，证明能有效沉默靶基因。本研究提供了一组具有潜在治疗应用的新型amiRNA，以及从内源性pri-miRNA生成和验证新型amiRNA的流程。
