一个工程T7RNA聚合酶,用于在mRNA合成中减少dsRNA副产品的高效协同转录封锁
Mathew Miller1, Oscar Alvizo1, Scott Baskerville1
1Codexis, Inc., 200 Penobscot Drive, Redwood City, CA 94063, USA. justin.dassie@codexis.com.
Faraday discussions
|June 4, 2024
概括
制造信使RNA (mRNA) 疗法是一项挑战. 一个新的酶,T7-68,改善了盖子模拟的结合,并减少了不必要的双链RNA (dsRNA) 副产品,以获得更可行的mRNA治疗方法.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 药物开发 药物开发
背景情况:
- 包括疫苗在内的信使RNA (mRNA) 疗法已经显示出显著的前景.
- 大规模的mRNA生产面临制造障碍,特别是低效的帽子模拟整合和在体外转录 (IVT) 过程中生成双链RNA (dsRNA) 副产品.
- 这些dsRNA副产品可以触发不良的免疫反应,并使净化复杂化.
研究的目的:
- 设计一种新型RNA聚合酶,能够进行高效的协同转录帽模拟整合.
- 在mRNA合成过程中减少不良dsRNA副产品的形成.
- 为了提高mRNA制造的可扩展性和成本效益.
主要方法:
- 一种新的RNA聚合酶的工程,指定T7-68.8.
- 使用T7-68.8评估二核和三核酸盖模拟结合的效率.
- 与野生型T7RNA聚合酶相比,用T7-68生成的IVT产品中的dSRNA含量量量化.
主要成果:
- 工程T7-68RNA聚合酶有效地结合了二核和三核酸盖类型,即使在较低度.
- 使用T7-68的IVT反应显示dSRNA副产品水平显著降低.
- 通过共同转录实现了高效率的模拟集成.
结论:
- 新型T7-68RNA聚合酶为mRNA疗法生产中的关键制造挑战提供了一个有前途的解决方案.
- 这种工程酶提高了帽子模拟合并的效率,并最大限度地降低了免疫性dsRNA副产品的形成.
- T7-68为治疗应用提供了更具可扩展性和潜在的成本效益的高质量的mRNA生产.
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