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在dsDNA上对多个mRNA片段进行单结合,促进区域修饰和翻译

Yunfan Xu¹, Shuopeng Qi¹, Gongrui Zhang¹

¹Key Laboratory of Bio-resource and Eco-environment of Ministry of Education, The College of Life Sciences, Sichuan University, 24 South Section 1, 1st Ring Road, Chengdu, Sichuan 610064, P.R. China.

Nucleic acids research

|January 8, 2025

概括

一种新方法使用双链DNA (dsDNA) 片来有效合成区域修饰的信使RNA (mRNA),改进mRNA疗法和疫苗.

科学领域:

分子生物学分子生物学
生物化学生物化学
在RNA治疗方面,RNA疗法.

背景情况:

特定区域的RNA修改对于mRNA研究,疫苗和免疫治疗至关重要.
目前使用单链DNA (ssDNA) 片的方法,由于RNA自我折叠,难以结长的mRNA片段.
合成修饰mRNA的挑战阻碍了治疗的发展.

研究的目的:

开发一种有效的生产区域修饰信使RNA (mRNA) 的战略.
克服现有方法在长和结构化的mRNA片段的结合方面的局限性.
研究特定RNA修改对翻译效率和免疫性的影响.

主要方法:

开发了一种新的策略,使用变质的长双链DNA (dsDNA) 作为结合 splint.
利用 dsDNA 的部分作为模板,用于 mRNA 片段的 in situ 转录.
合成的mRNA与N1-methylpseudouridine (m1ψ) 和5-methylcytidine (m5C) 修饰在特定区域使用一方法.

主要成果:

这种dsDNA splint策略显著提高了结合效率 (高达ssDNA的106倍).
变质的dsDNA形成了与mRNA片段的混合双重组,防止自我折叠.
与完全修改的mRNA相比,区域修改的mRNA显示了增强的翻译效率和降低的免疫性.

结论:

这种新的dsDNA切片策略提供了一种广泛适用的方法,用于生产区域修饰的mRNA.
这种方法增强了复杂的mRNA结构的合成,推进了mRNA治疗.
特定的修改,如3'UTR m1ψ,可以提高翻译效率和降低免疫性,为改进的mRNA基疗法铺平道路.

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