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在模板导向的mRNA合成过程中去除双链RNA污染物

S V Sharabrin¹, A A Bondar², E V Starostina³

¹State Research Center of Virology and Biotechnology "VECTOR", Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Koltsovo, Novosibirsk region, Russia. sharabrin.sv@gmail.com.

Bulletin of experimental biology and medicine

|June 19, 2024

概括

净化信使RNA (mRNA) 清除有害的双链RNA (dsRNA) 污染物. 这种新的基于纤维素的方法通过消除dsRNA杂质,确保了更安全的mRNA用于研究和潜在的疗法.

科学领域:

分子生物学分子生物学
生物技术是生物技术.
免疫学免疫学免疫学

背景情况:

双链RNA (dsRNA) 是体外合成的信使RNA (mRNA) 中常见的污染物.
这些dsRNA杂质是由于mRNA合成过程中T7RNA聚合酶的副作用活动引起的.
dsRNA污染物可以触发不必要的免疫反应,限制mRNA应用.

研究的目的:

开发和验证一种有效的方法,从mRNA中去除dSRNA污染物.
评估使用新方法净化mRNA的生物安全性.

主要方法:

采用了经过修改的mRNA净化技术,利用dsRNA在含有乙醇的缓冲器中选择性结合纤维素.
纯化的mRNA在试验室和活体中在小鼠模型中进行了测试.
测量了免疫反应标志物,包括CD69激活和干扰素α (IFNα) 释放.

主要成果:

在小鼠中,纤维素净化的mRNA没有显示淋巴细胞炎症标志物CD69的激活.
在用纤维素净化mRNA治疗的小鼠中没有检测到IFNα的增加释放.
净化方法有效地去除了dSRNA杂质,抗体检测证实了这一点.

结论:

开发的基于纤维素的净化方法有效地从mRNA中去除dSRNA污染物.
纯化的mRNA在生物学上是惰性的,在体内没有免疫激活的迹象.
这种方法为生产更安全的mRNA用于研究和治疗开发提供了有希望的解决方案.

关键词:

dsRNARNA 是一个它们是mRNARNA.在mRNA净化过程中.

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