具有稳定结构的工程化mRNA可最小化双链RNA形成并提高蛋白质表达
Engineered mRNAs With Stable Structures Minimize Double-stranded RNA Formation and Increase Protein Expression
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影响因子:4.5
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
DOI:
10.1016/j.jmb.2024.168822
摘要
合成信使RNA（mRNA）的治疗应用已在COVID-19疫苗中得到验证，并展现出在开发传染病和肿瘤疫苗方面的巨大潜力。然而，在体外转录（IVT）过程中产生的双链RNA（dsRNA）副产物可能降低mRNA治疗的效率并引发先天免疫反应。现有的去除dsRNA副产物的方法通常繁琐且劳动密集。在本研究中，我们发现mRNA的次级结构较松散以及序列中未配对的U碱基较多，通常会导致在IVT过程中生成更多的dsRNA副产物。我们进一步基于序列特性开发了预测dsRNA副产物形成的模型，以指导mRNA序列的优化，帮助最小化不必要的免疫反应并提高mRNA产物的蛋白表达。总体而言，本研究为开发具有较少dsRNA副产物和更高蛋白表达的高效mRNA治疗剂提供了新思路和方法学。
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.