具有稳定结构的工程mRNA最小化双链RNA的形成并增加蛋白质表达
Engineered mRNAs With Stable Structures Minimize Double-stranded RNA Formation and Increase Protein Expression
影响因子:4.50000
分区:生物学2区 / 生化与分子生物学3区
发表日期:2024 Nov 15
作者:
Qianshan Qin, Huayuan Yan, Weixiang Gao, Ruyin Cao, Guopeng Liu, Xiaojing Zhang, Niangang Wang, Wenjie Zuo, Lei Yuan, Peng Gao, Qi Liu
摘要
合成信息RNA（mRNA）的治疗用途已在Covid-19-19疫苗中得到验证，并在开发感染性和肿瘤疫苗中显示出巨大的潜力。然而，在体外转录过程（IVT）过程中产生的双链RNA（DSRNA）副产物可以降低基于mRNA的疗法的疗效并引起先天免疫反应。消除DSRNA副产品的现有方法通常是繁琐且劳动力密集的。在这项研究中，我们揭示了该序列中的松散mRNA二级结构和更多未配对的U碱基通常会导致在IVT过程中形成更多的DsRNA副产物。我们进一步开发了基于序列特征的DSRNA副产物形成的预测模型，以指导mRNA序列的优化，有助于最大程度地减少不需要的免疫反应并改善mRNA产物的蛋白质表达。总体而言，我们的研究提供了新的线索和方法，用于开发有效的mRNA疗法，并最小化dsRNA副产品并增加蛋白质的表达。
Abstract
The therapeutic use of synthetic message RNA (mRNA) has been validated in COVID-19 vaccines and shows enormous potential in developing infectious and oncological vaccines. However, double-stranded RNA (dsRNA) byproducts generated during the in vitro transcription (IVT) process can diminish the efficacy of mRNA-based therapeutics and provoke innate immune responses. Existing methods to eliminate dsRNA byproducts are often cumbersome and labor-intensive. In this study, we revealed that a loose mRNA secondary structure and more unpaired U bases in the sequence generally lead to the formation of more dsRNA byproducts during the IVT process. We further developed a predictive model for dsRNA byproducts formation based on sequence characteristics to guide the optimization of mRNA sequences, helping to minimize unwanted immune response and improve the protein expression of mRNA products. Collectively, our study provides novel clues and methodologies for developing effective mRNA therapeutics with minimized dsRNA byproducts and increased protein expression.