定向演化T7RNA聚合酶最小化dSRNA副产品,并使高保真性mRNA合成为苛刻的治疗应用
Weitong Qin1, Ting Nie1, Mohan Hei1
1State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
Research (Washington, D.C.)
|February 27, 2026
概括
工程T7RNA聚合酶 (T7RNAP) M30在mRNA合成中减少双链RNA副产品. 这种M30变体改善了治疗mRNA的产生,具有更高的效率和更低的免疫性.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 酶工程是什么? 酶工程是什么?
背景情况:
- T7RNA聚合酶 (T7RNAP) 对于治疗性信使RNA (mRNA) 合成至关重要.
- T7 RNAP转录的mRNA通常含有双链RNA (dsRNA) 副产品,触发免疫反应并阻碍净化.
- 减少dsRNA副产品对于改善mRNA质量和治疗应用至关重要.
研究的目的:
- 设计一种T7RNA聚合酶变体,提高催化效率并减少dsRNA副产品的形成.
- 评估工程变异的性能,以生产高质量的治疗mRNA.
- 调查观察到的改善的生物物理和结构基础.
主要方法:
- 在四轮的T7RNA聚合酶的定向演变中,使用基于aptamer的光激活滴滴分类系统.
- 工程变体 (M30) 对于催化效率,热稳定性和dSRNA副产品形成的表征.
- 在细胞和动物模型中评估由M30合成的mRNA用于蛋白质表达,免疫性和净化.
主要成果:
- 与37°C的野生类型T7RNAP相比,工程M30变体的催化效率增加了10倍.
- M30表现出增强的热稳定性和大约10倍低的dsRNA副产品的生产.
- 使用M30生产的mRNA导致人类细胞和小鼠的高效蛋白质表达,免疫性降低.
结论:
- M30 T7 RNAP变体是生产高质量的治疗mRNA的有希望的催化剂.
- M30的改进的DNA模板结合和减少的RNA结合亲缘关系有助于其增强的性能.
- 这种工程酶为克服当前mRNA合成技术的局限性提供了潜在的解决方案.
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