Li Ting, Liu Gangfeng, Bu Guolong, Xu Yien, He Caiyun, Zhao Gexin
State Key Laboratory of Oncology in South China, Guangdong Key Laboratory of Nasopharyngeal Carcinoma Diagnosis and Therapy, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou 510060, China.
Department of Oncology, The Affiliated Hospital of Qingdao Binhai University, 689 Haiya Road, Qingdao 266404, China.
Gene. 2025 Mar 20;942:149254. doi: 10.1016/j.gene.2025.149254. Epub 2025 Jan 15.
Advances in molecular medicine and biotechnology have demonstrated messenger RNA (mRNA)-based therapies to be a promising therapeutic modality for infectious diseases, genetic disorders, and cancers. However, key challenges persist, including low translation efficiency and short half-life of exogenous mRNA. The untranslated regions (UTRs) influence important parameters like mRNA stability and translation efficiency. This study adopted a combinatorial screening strategy to enhance exogenous mRNA translation efficiency by de novo designing 5'UTRs and combining them with multiple potential 3'UTRs. We designed a novel 5'UTR, 5UTR05, which exhibited comparable protein expression levels to the reference mRNA-1273 5'UTR that has been found to exhibit high expression in the COVID-19 vaccine development. Furthermore, the screening experiments found that combining 5UTR05 with both the 3'UTR of immunoglobulin heavy constant gamma 2 (IGHG2) and the 3'UTR of mitochondrially encoded 12S ribosomal RNA (mtRNR1) significantly improved mRNA translation efficiency, compared to individual 3'UTRs. Collectively, these findings provide valuable insights for UTR optimization strategies aimed at augmenting exogenous mRNA therapeutic translation. Continuing exploration of synergistic UTR combinations offers promise to advance customized mRNA constructs with optimized expression profiles tailored for diverse applications.
分子医学和生物技术的进展已证明基于信使核糖核酸（mRNA）的疗法是治疗传染病、遗传疾病和癌症的一种有前景的治疗方式。然而，关键挑战依然存在，包括外源mRNA的翻译效率低和半衰期短。非翻译区（UTR）会影响mRNA稳定性和翻译效率等重要参数。本研究采用组合筛选策略，通过从头设计5'UTR并将其与多个潜在的3'UTR相结合，来提高外源mRNA的翻译效率。我们设计了一种新型5'UTR，即5UTR05，其蛋白表达水平与在新冠疫苗研发中已发现具有高表达的参考mRNA-1273 5'UTR相当。此外，筛选实验发现，与单个3'UTR相比，将5UTR05与免疫球蛋白重链恒定γ2（IGHG2）的3'UTR和线粒体编码的12S核糖体RNA（mtRNR1）的3'UTR相结合，可显著提高mRNA的翻译效率。总体而言，这些发现为旨在增强外源mRNA治疗性翻译的UTR优化策略提供了有价值的见解。持续探索协同UTR组合有望推进定制的mRNA构建体，使其具有针对不同应用优化的表达谱。
