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通过端到端的自我准和拼接反应,使用Tetrahymena I组内 ribozyme进行高效的循环RNA工程

Kyung Hyun Lee¹, Seongcheol Kim¹, Jaehwi Song¹

¹R&D Center, Rznomics Inc, Seongnam 13486, Republic of Korea.

Molecular therapy. Nucleic acids

|August 28, 2023

概括

科学家们开发了一种新的体外试验方法,利用自我准和拼接反应来设计循环RNA (circRNA). 这种技术产生纯粹的circRNA,避免免疫反应,为疫苗和治疗开发提供了一个有希望的替代方案.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
在RNA治疗方面,RNA疗法.

背景情况:

循环RNA (circRNA) 在疫苗和治疗应用中比线性mRNA具有优势.
由于其潜力,circRNA的工程方法正在引起人们的注意.
现有的方法,如变的内子-外子 (PIE),可以引入不需要的碎片.

研究的目的:

开发一种新的体外方法,用于工程 circRNA.
为了利用端到端的自我准和拼接 (STS) 反应用于circRNA生成.
评估STS方法产生的circRNA的疗效和特征.

主要方法:

通过STS反应开发了一种体外circRNA工程方法,使用Tetrahymena I组内 ribozyme.
研究了P1螺旋结构在circRNA生成I组内中的作用.
将STS反应的有效性与变的内子-外子 (PIE) 反应进行比较.
使用离子对逆相高压液态染色学净化生成的circRNA.
进行了基于细胞的分析来评估circRNA的性能.

主要成果:

组I内子的P1螺旋结构足以通过STS反应生成circRNA.
在circRNA生成中,STS反应的疗效与PIE反应相当.
与PIE反应不同,STS反应避免了外来碎片 (内部痕),可能会减少先天免疫反应.
生成的circRNA得到了高效的净化,并在细胞中显示出高效的蛋白质表达和稳定性.
通过STS产生的circRNA表现出最小的先天免疫诱导.

结论:

一个新的 in vitro circRNA 工程策略使用端到端 STS 反应成功开发.
这种方法有效地产生了非常有用的circRNAs,为现有方法提供了替代方案.
没有内在痕使得这种circRNA策略在治疗应用中最大限度地减少不必要的免疫反应是有利的.

关键词:

MT:非编码RNAs的使用.替代RNA的RNA是一种替代RNA.对于RNA疗法来说,它是一种治疗方法.这是一种RNA疫苗.泰特拉希米纳是什么意思圆形的RNARNA是一个圆形的RNA.组I 内子组I 内子它们是mRNARNA.自己循环化的自我循环化.通过拼接的 ribozyme.

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